Difference between revisions of "Team:Kyoto/Notebook"

Line 168: Line 168:
 
     <div id="contents-wrapper">
 
     <div id="contents-wrapper">
 
    <div id="contents">
 
    <div id="contents">
 +
<!-- CONTENTS_HERE -->
  
 +
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0821" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/21</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>strain</td><td>Medium/ml</td></tr>
 +
<tr><td>8/20 transformation RBS</td><td>PG(AMP) /6</td></tr>
 +
<tr><td>8/20 dilution INP7</td><td>PG(CP) /6</td></tr>
 +
<tr><td>8/17 transformation 3c5, RFP</td><td>PG(CP) /6</td></tr>
 +
<tr><td>8/20 transformation p+ [1]</td><td>PG(AMP) /6</td></tr>
 +
<tr><td>8/20 transformation p- [1]</td><td>PG(AMP) /6</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0821" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/21</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /1.5</td><td></td></tr>
 +
<tr><td>2</td><td>8/21 cloning PCR scfv[1] /2</td><td></td></tr>
 +
<tr><td>3</td><td>8/21 cloning PCR scfv[2] /2</td><td></td></tr>
 +
<tr><td>4</td><td>nega /2</td><td></td></tr>
 +
<tr><td>5</td><td>8/21 colony PCR p+ [1] /2</td><td></td></tr>
 +
<tr><td>6</td><td>8/21 colony PCR p+ [2] /2</td><td></td></tr>
 +
<tr><td>7</td><td>8/21 colony PCR p+ [3] /2</td><td></td></tr>
 +
<tr><td>8</td><td>8/21 colony PCR p- [1] /2</td><td></td></tr>
 +
<tr><td>9</td><td>8/21 colony PCR p- [2] /2</td><td></td></tr>
 +
<tr><td>10</td><td>8/21 colony PCR p- [3] /2</td><td></td></tr>
 +
<tr><td>11</td><td>8/21 colony PCR parts p+ /2</td><td></td></tr>
 +
<tr><td>12</td><td>8/21 colony PCR parts p- /2</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/9/9d/T--Kyoto--8_21_No34_Electrophoresis_Kaneko_05_20_05_13_01.jpg">
 +
</div>
 +
<p class="keln_note">[img35]<br>2, 3, 4 cloning PCR failed.<br>1 forgot to put ladder<br>5, 7, 8, 9 strong band<br>6 weak<br>10 failed<br>P+[1] and p-[1] -> liquid culture</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0824" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/24</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>sample/&microl</th><th>MilliQ (&microl)</th><th>Buffer /&microl</th><th>Ligase(&microl)</th><th>Total(&microl)</th></tr>
 +
<tr><td>8/19 mutation PCR scfv /5</td><td>3.5</td><td>1</td><td>0.5</td><td>10</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0824" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/24</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>8/20 transformation m.scfv colonyA /6</td><td>PG(AMP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0824" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/24</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>8/20 Liquid culture RBS</td><td>124.5</td><td>1.65</td><td>1.02</td></tr>
 +
<tr><td>8/20 Liquid culture INP</td><td>147.8</td><td>1.66</td><td>0.98</td></tr>
 +
<tr><td>8/20 Liquid culture 3c5.RFP</td><td>234.8</td><td>1.79</td><td>1.62</td></tr>
 +
<tr><td>8/20 Liquid culture P+[1]</td><td>40.3</td><td>1.78</td><td>1.60</td></tr>
 +
<tr><td>8/20 Liquid culture P-[1]</td><td>56.7</td><td>1.80</td><td>2.05</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Iguchi -->
 +
<div class="keln_container">
 +
<a name="0825" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/25</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Li, Uchino, Notsu, Kaneko</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>8/24 LIquid Culture m.scfv colonyA</td><td>-</td><td>-</td><td>-</td></tr>
 +
<tr><td>8/24 LIquid Culture m.scfv colonyB</td><td>-</td><td>-</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0825" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/25</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher"></span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /1.5</td><td></td></tr>
 +
<tr><td>2</td><td>8/24 miniprep p+(J61002) /2</td><td></td></tr>
 +
<tr><td>3</td><td>8/24 miniprep p-(J61002) /2</td><td></td></tr>
 +
<tr><td>4</td><td>8/24 miniprep RFP (3c5) /2</td><td></td></tr>
 +
<tr><td>5</td><td>8/24 miniprep INP (1c3) /2</td><td></td></tr>
 +
<tr><td>6</td><td>8/24 miniprep RBS (1A2) /2</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/f/fd/T--Kyoto--8_25_No38_Electrophoresis_Kaneko_05_20_05_22_35.jpg">
 +
</div>
 +
<p class="keln_note">[img38]</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0825" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/25</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td></td><td>p+/24.8</td><td></td><td></td><td>1</td><td>1</td><td>10xH /4</td><td></td><td>9.2</td><td>40</td></tr>
 +
<tr><td></td><td>IINP /6.8</td><td></td><td>1</td><td></td><td>1</td><td>10xM /4</td><td></td><td>27.2</td><td>40</td></tr>
 +
<tr><td></td><td>p- /17.6</td><td></td><td></td><td>1</td><td>1</td><td>10xH /4</td><td></td><td>16.4</td><td>40</td></tr>
 +
<tr><td></td><td>RBS /8.0</td><td></td><td></td><td>1</td><td>1</td><td>10xH /4</td><td></td><td>26</td><td>40</td></tr>
 +
<tr><td></td><td>RFP /4.3</td><td>1</td><td></td><td></td><td>1</td><td>10xH /4</td><td></td><td>29.7</td><td>40</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0825" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/25</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Uchino, Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder </td><td>6</td></tr>
 +
<tr><td>2, 3</td><td>8/25 digestion p+</td><td>24 each</td></tr>
 +
<tr><td>4, 5</td><td>8/25 digestion p- </td><td>24 each</td></tr>
 +
<tr><td>6, 7</td><td>8/25 digestion RFP </td><td>24 each</td></tr>
 +
<tr><td>8, 9</td><td>8/25 digestion INP </td><td>24 each</td></tr>
 +
<tr><td>10, 11</td><td>8/25 digestion RBS </td><td>24 each</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/7/77/T--Kyoto--8_25_No40_Gel_Extraction_Kaneko_05_20_05_28_41.JPG">
 +
</div>
 +
<p class="keln_note">[img40]</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0825" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/25</h3></span>
 +
<span class="keln_exp"><h4>Actvation check of Xba and Pst</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>INP(&microl)</th><th>Xba(&microl)</th><th>Pst(&microl)</th><th>10xM(&microl)</th><th>BSA(&microl)</th><th>MilliQ(&microl)</th><th>Total(&microl)</th></tr>
 +
<tr><td>0.68</td><td>0.1</td><td>0.1</td><td>1</td><td>0.2</td><td>7.92</td><td>10</td></tr>
 +
<tr><td>0.68</td><td>0.1</td><td>-</td><td>1</td><td>0.2</td><td>8.02</td><td>10</td></tr>
 +
<tr><td>0.68</td><td>-</td><td>0.1</td><td>1</td><td>0.2</td><td>8.02</td><td>10</td></tr>
 +
<tr><td>0.68</td><td>-</td><td>-</td><td>1</td><td>0.2</td><td>8.12</td><td>10</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0825" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/25</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /3</td><td></td></tr>
 +
<tr><td>2</td><td>[1] (inp xp cut) /10</td><td></td></tr>
 +
<tr><td>3</td><td>[2] (inp x cut) /10</td><td></td></tr>
 +
<tr><td>4</td><td>[3] (inp p cut) /10</td><td></td></tr>
 +
<tr><td>5</td><td>[4] (inp no cut) /10</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/4/46/T--Kyoto--8_25_No41_2_Electrophoresis_Kaneko_05_20_05_34_47.jpg">
 +
</div>
 +
<p class="keln_note">[img42]</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0826" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/26</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Uchino, Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td></td><td>8/24 miniprep INP 1c3 /0.68</td><td>-</td><td>1.0</td><td></td><td></td><td></td><td>0.2</td><td>7.12</td><td>10</td></tr>
 +
<tr><td></td><td>8/15 miniprep RFP psb1c3 /1.3</td><td>-</td><td>1.0</td><td></td><td></td><td></td><td>0.2</td><td>6.5</td><td>10</td></tr>
 +
<tr><td></td><td>8/24 miniprep INP 1c3 /0.68</td><td>0.1</td><td>1.0</td><td></td><td></td><td></td><td>0.2</td><td>7.02</td><td>10</td></tr>
 +
<tr><td></td><td>8/15 miniprep RFP psb1c3 /1.3</td><td>0.1</td><td>1.0</td><td></td><td></td><td></td><td>0.2</td><td>6.4</td><td>10</td></tr>
 +
<tr><td></td><td>8/24 miniprep INP 1c3/ 0.68</td><td>0.1</td><td>-</td><td></td><td></td><td></td><td>0.2</td><td>8.02</td><td>10</td></tr>
 +
<tr><td></td><td>8/15 miniprep RFP psb1c3 /1.3</td><td>0.1</td><td>-</td><td></td><td></td><td></td><td>0.2</td><td>7.4</td><td>10</td></tr>
 +
<tr><td></td><td>8/24 miniprep INP 1c3 /0.68</td><td>-</td><td>-</td><td></td><td></td><td></td><td>0.2</td><td>8.12</td><td>10</td></tr>
 +
<tr><td></td><td>8/15 miniprep RFP psb1c3 /1.3</td><td>-</td><td>-</td><td></td><td></td><td></td><td>0.2</td><td>7.5</td><td>10</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0826" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/26</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>scfv from MK</td><td>c.scfv fw/0.75</td><td>c.scfv rv/0.75</td><td>2x KAPA Hifi /12.5</td><td>11</td><td></td><td>25</td></tr>
 +
<tr><td>none</td><td>c.scfv fw/0.75</td><td>c.scfv rv/0.75</td><td>2x KAPA Hifi /5</td><td>4.4</td><td></td><td>10</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0826" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/26</h3></span>
 +
<span class="keln_exp"><h4>Activation Check of Xba</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder/2</td><td></td></tr>
 +
<tr><td>2</td><td>[8] RFP/10</td><td></td></tr>
 +
<tr><td>3</td><td>[6] RFP EcoRI/10</td><td></td></tr>
 +
<tr><td>4</td><td>[2] RFP XbaI/10</td><td></td></tr>
 +
<tr><td>5</td><td>[4] RFP EcoRI, XbaI /10</td><td></td></tr>
 +
<tr><td>6</td><td>[7] INP /10</td><td></td></tr>
 +
<tr><td>7</td><td>[5] INP EcoRI /10</td><td></td></tr>
 +
<tr><td>8</td><td>[1] INP XbaI /10</td><td></td></tr>
 +
<tr><td>9</td><td>[3] INP EcoRI, XbaI/10</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/1/17/T--Kyoto--8_26_No45_Electrophoresis_Kaneko_05_20_05_42_54.jpg">
 +
</div>
 +
<p class="keln_note">[img45-1]<br>[img45-2]</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Iguchi -->
 +
<div class="keln_container">
 +
<a name="0827" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/27</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>2</td><td>1kb ladder</td><td>2</td></tr>
 +
<tr><td>3</td><td>8/25 PCR scfv</td><td>5</td></tr>
 +
<tr><td>4</td><td>8/25 PCR none</td><td>5</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/f/fc/T--Kyoto--8_27_No46_Electrophoresis_Iguchi_10_17_06_08_50.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Iguchi -->
 +
<div class="keln_container">
 +
<a name="0827" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/27</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Predenature</td><td>95</td><td>3min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>98</td><td>20sec</td><td>25</td></tr>
 +
<tr><td>Annealing</td><td>65</td><td>15sec</td><td>25</td></tr>
 +
<tr><td>Extension</td><td>72</td><td>30sec</td><td>25</td></tr>
 +
<tr><td>Final Elogation</td><td>72</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>4</td><td>-</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Iguchi -->
 +
<div class="keln_container">
 +
<a name="0827" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/27</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>s.BAN</td><td>BAN.Fw/ 0.75</td><td>BAN.Rv/ 0.75</td><td></td><td>11</td><td>2xKAPA HiFi/ 12.5</td><td>25</td></tr>
 +
<tr><td>s.BAN</td><td>BAN.Fw/ 0.75</td><td>BAN.Rv/ 0.75</td><td></td><td>11</td><td>2xKAPA HiFi/ 12.5</td><td>25</td></tr>
 +
<tr><td>s.BAN</td><td>BAN.Fw/ 0.75</td><td>BAN.Histag/ 0.75</td><td></td><td>11</td><td>2xKAPA HiFi/ 12.5</td><td>25</td></tr>
 +
<tr><td>scfv</td><td>scfv.Fw/ 0.75</td><td>scfv.Rv/ 0.75</td><td></td><td>11</td><td>2xKAPA HiFi/ 12.5</td><td>25</td></tr>
 +
<tr><td>8/24 miniprep INP</td><td>INP.Fw/ 0.75</td><td>INP.Rv/ 0.75</td><td></td><td>11</td><td>2xKAPA HiFi/ 12.5</td><td>25</td></tr>
 +
<tr><td>8/24 miniprep INP</td><td>INP.Fw/ 0.75</td><td>INP.Rv.Histag/ 0.75</td><td></td><td>11</td><td>2xKAPA HiFi/ 12.5</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0814" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/14</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Li, Uchino, Kaneko</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>RFP 0.5ng/&microl</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>RFP 5ng/&microl</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>RFP 10ng/&microl</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>RFP 20ng/&microl</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>RFP 50ng/&microl</td><td>2</td><td>DH5α /20</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0814" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/14</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Li, Uchino, Kaneko</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>10/1 PQ(3)</td><td>about 23.5&microl</td></tr>
 +
<tr><td>2</td><td>1kb ladder</td><td>6</td></tr>
 +
<tr><td>3</td><td>10/1 PQ(3)</td><td>about 23.5&microl</td></tr>
 +
<tr><td>4</td><td>10/1 PQ(3)</td><td>about 24&microl</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/7/7a/T--Kyoto--8_14_No1_Gel_Extraction_Notsu_09_19_12_14_09No.20814.jpg">
 +
</div>
 +
<p class="keln_note">Total volume of samples of lane 1~3 was 71.5&microl and it was divided into three.</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Egashira -->
 +
<div class="keln_container">
 +
<a name="0814" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/14</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Li, Uchino, Kaneko</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>3C5</td><td>10&microl</td><td>DH5留 20&microl</td><td>LB(CP)</td></tr>
 +
<tr><td>P+</td><td>10&microl</td><td>DH5留 20&microl</td><td>LB(CP)</td></tr>
 +
<tr><td>P-</td><td>10&microl</td><td>DH5留 20&microl</td><td>LB(CP)</td></tr>
 +
<tr><td>RBS</td><td>10&microl</td><td>DH5留 20&microl</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0814" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/14</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Li, Uchino, Iguchi, Kaneko</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>8/13 transformation RFP 20ng/&microl colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>8/13 transformation part RFP colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>8/13 transformation part RFP colony2</td><td>LB(CP)</td></tr>
 +
<tr><td>8/13 transformation part RFP colony3</td><td>LB(CP)</td></tr>
 +
<tr><td>8/13 transformation part RFP colony4</td><td>LB(CP)</td></tr>
 +
<tr><td>8/13 transformation part RFP colony5</td><td>LB(CP)</td></tr>
 +
<tr><td>8/13 transformation part RFP colony6</td><td>LB(CP)</td></tr>
 +
<tr><td>8/13 transformation part RFP colony7</td><td>LB(CP)</td></tr>
 +
<tr><td>8/13 transformation part RFP colony8</td><td>LB(CP)</td></tr>
 +
<tr><td>8/13 transformation part RFP colony9</td><td>LB(CP)</td></tr>
 +
<tr><td>8/13 transformation part RFP colony10</td><td>LB(CP)</td></tr>
 +
<tr><td></td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0815" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/15</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Li, Uchino, Notsu, Kaneko, Watanabe</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>RFP1 /2&microl</td><td>82.5</td><td>1.40</td><td>1.22</td></tr>
 +
<tr><td>RFP2 /4&microl</td><td>142.5</td><td>1.69</td><td>2.05</td></tr>
 +
<tr><td>RFP3 /4&microl</td><td>50</td><td>1.55</td><td>1.71</td></tr>
 +
<tr><td>RFP4 /4&microl</td><td>57.5</td><td>1.64</td><td>1.88</td></tr>
 +
<tr><td>RFP6 /4&microl</td><td>98.75</td><td>1.71</td><td>1.40</td></tr>
 +
<tr><td>RFP(20) /4&microl</td><td>76.25</td><td>1.67</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Egashira -->
 +
<div class="keln_container">
 +
<a name="0815" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/15</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Notsu, Kaneko</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Products</th></tr>
 +
<tr><td>1</td><td>1kb ladder 3&microl</td></tr>
 +
<tr><td>2</td><td>RFP1 10&microl</td></tr>
 +
<tr><td>3</td><td>RFP2 10&microl</td></tr>
 +
<tr><td>4</td><td>RFP3 10&microl</td></tr>
 +
<tr><td>5</td><td>RFP4 10&microl</td></tr>
 +
<tr><td>6</td><td>RFP6 10&microl</td></tr>
 +
<tr><td>7</td><td>RFP(20) 10&microl</td></tr>
 +
<tr><td>8</td><td>PCR1 5&microl</td></tr>
 +
<tr><td>9</td><td>PCR2 5&microl</td></tr>
 +
<tr><td>10</td><td>PCR3 5&microl</td></tr>
 +
<tr><td>11</td><td>PCR4 5&microl</td></tr>
 +
<tr><td>12</td><td> PCR5 5 &microl</td></tr>
 +
<tr><td>13</td><td>1kb ladder 3&microl</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/5/5e/T--Kyoto--8_15_No6_Electrophoresis_Egashira_02_08_02_44_07.JPG">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Egashira -->
 +
<div class="keln_container">
 +
<a name="0815" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/15</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Uchino, Notsu, Kaneko</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>8/15Miniprep PSB1C3-RFP1</td><td>24</td><td>1</td><td>0</td><td>0</td><td>1</td><td>5(H)</td><td>0</td><td>19</td><td>50</td></tr>
 +
<tr><td>8/15Miniprep PSB1C3-RFP2</td><td>14</td><td>1</td><td>0</td><td>0</td><td>1</td><td>5(H)</td><td>0</td><td>29</td><td>50</td></tr>
 +
<tr><td>8/15Miniprep PSB1C3-RFP3</td><td>40</td><td>1</td><td>0</td><td>0</td><td>1</td><td>5(H)</td><td>0</td><td>3</td><td>50</td></tr>
 +
<tr><td>8/15Miniprep PSB1C3-RFP4</td><td>35</td><td>1</td><td>0</td><td>0</td><td>1</td><td>5(H)</td><td>0</td><td>8</td><td>50</td></tr>
 +
<tr><td>8/15Miniprep PSB1C3-RFP6</td><td>20</td><td>1</td><td>0</td><td>0</td><td>1</td><td>5(H)</td><td>0</td><td>23</td><td>50</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0815" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/15</h3></span>
 +
<span class="keln_exp"><h4>colony PCR</h4></span>
 +
<span class="keln_researcher">Uchino, Notsu, Kaneko, Watanabe</span>
 +
<table class="keln_table">
 +
<tr><th>colony                                                                        Primers</th></tr>
 +
<tr><td>8/14 transformation pSB1C3-RFP1                    VF+VR</td></tr>
 +
<tr><td>8/14 transformation pSB1C3-RFP2                    VF+VR</td></tr>
 +
<tr><td>8/14 transformation pSB1C3-RFP3                    VF+VR</td></tr>
 +
<tr><td>8/14 transformation pSB1C3-RFP4                    VF+VR</td></tr>
 +
<tr><td>8/14 transformation pSB1C3-RFP1                    VF+VR</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0815" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/15</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Uchino, Notsu, Kaneko, Watanabe</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Pre Denature</td><td>94℃</td><td>2min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>94℃</td><td>30sec</td><td>30</td></tr>
 +
<tr><td>Annealing</td><td>55℃</td><td>30sec</td><td>30</td></tr>
 +
<tr><td>Extension</td><td>68℃</td><td>1min</td><td>30</td></tr>
 +
<tr><td>Final elongation</td><td>4℃</td><td>7min</td><td></td></tr>
 +
<tr><td>Final Holding</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0816" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/16</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Li, Iguchi, Notsu, Kaneko, Watanabe</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td>10</td></tr>
 +
<tr><td>2</td><td>blank</td><td></td></tr>
 +
<tr><td>3</td><td>8/15 digestion pSB1C3-RFP 1</td><td>55</td></tr>
 +
<tr><td>4</td><td>8/15 digestion pSB1C3-RFP 2</td><td>55</td></tr>
 +
<tr><td>5</td><td>8/15 digestion pSB1C3-RFP 3</td><td>36</td></tr>
 +
<tr><td>6</td><td>8/15 digestion pSB1C3-RFP 4</td><td>36</td></tr>
 +
<tr><td>7</td><td>8/15 digestion pSB1C3-RFP 6</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/7/77/T--Kyoto--8_16_No10_Gel_Extraction_Notsu_09_26_01_32_24.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0816" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/16</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Li, Iguchi, Notsu, Kaneko, Watanabe</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>8/16 Gel Extraction RFP1 /1</td><td></td></tr>
 +
<tr><td>2</td><td>8/16 Gel Extraction RFP2 /1</td><td></td></tr>
 +
<tr><td>3</td><td>8/16 Gel Extraction RFP3 /1</td><td></td></tr>
 +
<tr><td>4</td><td>8/16 Gel Extraction RFP4 /1</td><td></td></tr>
 +
<tr><td>5</td><td>blank</td><td></td></tr>
 +
<tr><td>6</td><td>8/16 pSB1C3-vector /1</td><td></td></tr>
 +
<tr><td>7</td><td>8/16 pSB1C3-vector /1</td><td></td></tr>
 +
<tr><td>8</td><td></td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/6/6f/T--Kyoto--8_16_No11_Electrophoresis_Notsu_09_26_01_55_23.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0816" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/16</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Li, Iguchi, Notsu, Kaneko, Watanabe</span>
 +
<table class="keln_table">
 +
<tr><th>Vector DNA /(&microl)                  insert DNA /(&microl)                  ligase /(&microl)              10×buffer /(&microl)          MilliQ /(&microl)                Total /(&microl)                  </th></tr>
 +
<tr><td>pSB1C3 EPcut /1                    RFP /4                                    0.5                              1                              3                              9.5</td></tr>
 +
<tr><td>pSB1C3 EPcut /1                    RFP /2                                    0.5                              1                              5                              9.5</td></tr>
 +
<tr><td>pSB1C3 EPcut /1                    RFP /4                                    0.5                              1                              3                              9.5</td></tr>
 +
<tr><td>pSB1C3 EPcut /1                    RFP /4                                    0.5                              1                              3                              9.5</td></tr>
 +
<tr><td>pSB1C3 EPcut /1                        -                                          0.5                              1                              7                              9.5</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0817" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/17</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>8/16 transformation pSB3C5 RFP1</td><td>LB(CP) /6</td></tr>
 +
<tr><td>8/16 transformation pSB3C5 RFP2</td><td>LB(CP) /6</td></tr>
 +
<tr><td>8/16 transformation pSB3C5 RFP1</td><td>LB(CP) /6</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0817" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/17</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>pSB1C3 P+</td><td>2</td><td>DH5α /20</td><td>LB</td></tr>
 +
<tr><td>pSB1C3 P-</td><td>2</td><td>DH5α /20</td><td>LB</td></tr>
 +
<tr><td>pSB1C3 RBS</td><td>2</td><td>DH5α /20</td><td>LB</td></tr>
 +
<tr><td>pSB3C5 RFP</td><td>2</td><td>DH5α /20</td><td>LB</td></tr>
 +
<tr><td>8/16 ligation pSB1C3 RFP1</td><td>2</td><td>DH5α</td><td>LB</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0817" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/17</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Iguchi</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kbp ladder /3</td><td></td></tr>
 +
<tr><td>2</td><td>blank</td><td></td></tr>
 +
<tr><td>3</td><td>pSB1C3-RFP(ligate)1 /2</td><td></td></tr>
 +
<tr><td>4</td><td>pSB1C3-RFP(ligate)2 /2</td><td></td></tr>
 +
<tr><td>5</td><td>pSB1C3-RFP(ligate)3 /2</td><td></td></tr>
 +
<tr><td>6</td><td>pSB1C3-RFP(ligate)4 /3</td><td></td></tr>
 +
<tr><td>7</td><td>Vector (Ctrl) /5</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/c/c7/T--Kyoto--8_17_No14_Electrophoresis_Notsu_09_29_12_26_11.JPG">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0818" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/18</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Li, Iguchi, Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>INP [1] BBa_K811005</td><td>LB(CP) / 6ml</td></tr>
 +
<tr><td>INP [2] BBa_K811005</td><td>LB(CP) / 6ml</td></tr>
 +
<tr><td>INP [3] BBa_K811005</td><td>LB(CP) / 6ml</td></tr>
 +
<tr><td>scfv04 [1] BBa_K875004</td><td>LB(CP) / 6ml</td></tr>
 +
<tr><td>scfv04 [2] BBa_K875004</td><td>LB(CP) / 6ml</td></tr>
 +
<tr><td>scfv04 [3] BBa_K875004</td><td>LB(CP) / 6ml</td></tr>
 +
<tr><td>scfv06 [1] BBa__K875006</td><td>LB(CP) / 6ml</td></tr>
 +
<tr><td>scfv06 [2] BBa_K875006</td><td>LB(CP) / 6ml</td></tr>
 +
<tr><td>scfv06 [3] BBa_K875006</td><td>LB(CP) / 6ml</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0818" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/18</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Li, Iguchi, Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>8/17 Liquid Culture 3C5 [1] / 5&microl</td><td>77</td><td>1.69</td><td>1.45</td></tr>
 +
<tr><td>8/17 Liquid Culture 3C5 [2] / 2&microl</td><td>40</td><td>1.67</td><td>1.17</td></tr>
 +
<tr><td>8/17 Liquid Culture 3C5 [3] / 2&microl</td><td>15</td><td>3.75</td><td>3.70</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0818" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/18</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /1</td><td>-</td></tr>
 +
<tr><td>2</td><td>[1] psb3c5(8/18 miniprep) /4.2</td><td>3807</td></tr>
 +
<tr><td>3</td><td>[2] psb3c5(8/18 miniprep) /4.2</td><td>3807</td></tr>
 +
<tr><td>4</td><td>[1'] psb3c5(8/18 miniprep) /4.2</td><td>3807</td></tr>
 +
<tr><td>5</td><td>[2'] psb3c5(8/18 miniprep) /4.2</td><td>3807</td></tr>
 +
<tr><td>6</td><td>[3'] psb3c5(8/18 miniprep) /4.2</td><td>3807</td></tr>
 +
</table>
 +
<img src="">https://static.igem.org/mediawiki/2016/c/c5/T--Kyoto--8_18_No18_Electrophoresis_Kaneko_05_20_04_12_53.jpg
 +
</div>
 +
<p class="keln_note">[img18]</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0818" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/18</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Li, Iguchi, Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td></td><td>psb3c5(8/18miniprep) [1'], [2'], [3'] mixed / 50</td><td>1.0</td><td></td><td></td><td>1.0</td><td>5.6</td><td></td><td></td><td>56.6</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0819" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/19</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Li, Iguchi, Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>BBa_ J61002 p+</td><td>2</td><td>DH5a /20</td><td>LB(Amp)</td></tr>
 +
<tr><td>BBa_J61002 p-</td><td>2</td><td>DH5a /20</td><td>LB(Amp)</td></tr>
 +
<tr><td>BBa_1A2 RBS</td><td>2</td><td>DH5a /20</td><td>LB(Amp)</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">Failed due to deactivation of Amp</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0819" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/19</h3></span>
 +
<span class="keln_exp"><h4>MutationPCR</h4></span>
 +
<span class="keln_researcher"></span>
 +
<table class="keln_table">
 +
<tr><th>Templates /&microl</th><th>Primer /&microl</th><th>Primer STAR Max Premix /&microl</th><th>MilliQ /&microl</th><th>Total(&microl)</th></tr>
 +
<tr><td>scfv plasmid /0.1</td><td>m.scfv FW /1, m.scfvRV/1</td><td>25</td><td>22.9</td><td>50</td></tr>
 +
<tr><td>scfv plasmid (nega) /0.1</td><td>0</td><td>5</td><td>4.9</td><td>10</td></tr>
 +
<tr><td>(nega) /0</td><td>m.scfv FW/0.2, m.scfvRV/0.2</td><td>5</td><td>4.6</td><td>10</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">---30cycle---<br>98℃ 10''<br>55℃ 15''<br>72℃ 20''<br>----------------<br>4℃ holding</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0819" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/19</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder/2</td><td>-</td></tr>
 +
<tr><td>2</td><td>scfv plasmid +primer(m.scfv FW, m.scfv RV) mutation PCR product /5</td><td>3600</td></tr>
 +
<tr><td>3</td><td>scfv plasmid mutation PCR product /5</td><td>-</td></tr>
 +
<tr><td>4</td><td>primer(m.scfv FW, m.scfv RV) mutation PCR product /5</td><td>-</td></tr>
 +
<tr><td>5</td><td>psb3c5 EPcut product /5</td><td>2600(backborn), 1200(inseert<RFP>)</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/6/64/T--Kyoto--8_19_No22_Electrophoresis_Kaneko_05_20_04_27_06.jpg">
 +
</div>
 +
<p class="keln_note">[img22]</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0819" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/19</h3></span>
 +
<span class="keln_exp"><h4>Concentration</h4></span>
 +
<span class="keln_researcher">Li, Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>OD600</th><th>Bacterial cells /ml</th></tr>
 +
<tr><td>INP [1] K811005</td><td>0.643</td><td>5.14x10^8</td></tr>
 +
<tr><td>scfv06 K875006</td><td>0.634</td><td>5.07x10^8</td></tr>
 +
<tr><td>scfv04 K875004</td><td>0.618</td><td>4.94x10^8</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">1 OD 600 equals to 8x10^8 cells /ml</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0819" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/19</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Li, Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>scfv 04[1] K875004 / 3&microl</td><td>56</td><td>1.63</td><td>1.45</td></tr>
 +
<tr><td>scfv 04[2] K875004 / 3&microl</td><td></td><td></td><td></td></tr>
 +
<tr><td>scfv 04[3] K875004 / 3&microl</td><td></td><td></td><td></td></tr>
 +
<tr><td>scfv 06[1] K875006 /3&microl</td><td></td><td></td><td></td></tr>
 +
<tr><td>scfv 06[2] K875006 /3&microl</td><td></td><td></td><td></td></tr>
 +
<tr><td>scfv 06[3] K875006 /3&microl</td><td></td><td></td><td></td></tr>
 +
<tr><td>INP [1] K811005 /3&microl</td><td></td><td></td><td></td></tr>
 +
<tr><td>INP [2] K811005 /3&microl</td><td></td><td></td><td></td></tr>
 +
<tr><td>INP [3] K811005 /3&microl</td><td></td><td></td><td></td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">There was a problem with an instrument</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0820" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/20</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Li, Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>8/19 mutation PCR scfv plasmid (nega)</td><td>1</td><td>DH5a/ 20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">8/19 mutation PCR:<br>1&microl extraction of scfv plasmid (nega)/10&microl was folded 300 times.</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0820" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/20</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Li, Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>8/19 mutation PCR scfv plasmid</td><td>1 [note:1]</td><td>DH5a/20</td><td>LB(AMP)</td></tr>
 +
<tr><td>8/19 mutation PCR scfv plasmid (nega)</td><td>1 [note:2]</td><td>DH5a/20</td><td>LB(AMP)</td></tr>
 +
<tr><td>BBa_J61002 p+</td><td>1</td><td>DH5a/20</td><td>LB(AMP)</td></tr>
 +
<tr><td>BBa_J61002 p-</td><td>1</td><td>DH5a/20</td><td>LB(AMP)</td></tr>
 +
<tr><td>BBa_1A2 RBS</td><td>1</td><td>DH5a/20</td><td>LB(AMP)</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">[note:1]<br>8/19 mutation PCR:<br>1&microl extraction of scfv plasmid (nega)/50&microl was folded 300 times.<br>[note:2]<br>8/19 mutation PCR:<br>1&microl extraction of scfv plasmid (nega)/10&microl was folded 300 times.</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0820" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/20</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /1.5</td><td></td></tr>
 +
<tr><td>2</td><td>9/19 miniprep scfv04 [1]</td><td></td></tr>
 +
<tr><td>3</td><td>9/19 miniprep scfv04 [2]</td><td></td></tr>
 +
<tr><td>4</td><td>9/19 miniprep scfv04 [3]</td><td></td></tr>
 +
<tr><td>5</td><td>9/19 miniprep scfv06 [1]</td><td></td></tr>
 +
<tr><td>6</td><td>9/19 miniprep scfv06 [2]</td><td></td></tr>
 +
<tr><td>7</td><td>9/19 miniprep scfv06 [3]</td><td></td></tr>
 +
<tr><td>8</td><td>9/19 miniprep INP [1]</td><td></td></tr>
 +
<tr><td>9</td><td>9/19 miniprep INP [2]</td><td></td></tr>
 +
<tr><td>10</td><td>9/19 miniprep INP [3]</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/5/53/T--Kyoto--8_20_No26_Electrophoresis_Kaneko_05_20_04_44_45.jpg">
 +
</div>
 +
<p class="keln_note">[img26]</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0821" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/21</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Colony</th><th>Primers</th></tr>
 +
<tr><td>8/20 transformation RBS</td><td>VF+VR</td></tr>
 +
<tr><td>8/20 transformation p+</td><td>VF+VR</td></tr>
 +
<tr><td>8/20 transformation p-</td><td>VF+VR</td></tr>
 +
<tr><td>8/20 dilution INP</td><td>VF+VR</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0821" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/21</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Pre</td><td>94℃</td><td>2min</td><td>1</td></tr>
 +
<tr><td>De</td><td>94℃</td><td>30 sec</td><td>30</td></tr>
 +
<tr><td>An</td><td>55℃</td><td>30 sec</td><td>30</td></tr>
 +
<tr><td>Ex</td><td>68℃</td><td>1min</td><td>30</td></tr>
 +
<tr><td>Final E</td><td>4℃</td><td>7min</td><td>1</td></tr>
 +
<tr><td>Final Hold</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">Autoclaved distrilled water /23&microl<br>2xQuick taq /25&microl<br>10 pmol/&microl primer (VF) /1&microl<br>10 pmol/&microl primer (VR) /1&microl<br>Template DNA /0&microl<br>Total 50&microl</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0821" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/21</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /2</td><td></td></tr>
 +
<tr><td>2</td><td>8/21 colony PCR RBS /all</td><td></td></tr>
 +
<tr><td>3</td><td>8/21 colony PCR p+ /all</td><td></td></tr>
 +
<tr><td>4</td><td>8/21 colony PCR p- /all</td><td></td></tr>
 +
<tr><td>5</td><td>8/21 colony PCR INP /all</td><td></td></tr>
 +
<tr><td>6</td><td>8/21 colony PCR Nega /all</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/0/04/T--Kyoto--8_21_No28_Electrophoresis_Kaneko_05_20_04_54_01%2C1.jpg"><img src="https://static.igem.org/mediawiki/2016/b/b5/T--Kyoto--8_21_No28_Electrophoresis_Kaneko_05_20_04_54_01%2C2.jpg">
 +
</div>
 +
<p class="keln_note">[img29-1]<br>weak band except INP -> retry<br>colony PCR and Electrophoresis were conducted under the same condition.<br>[img29-2]<br>RBS and INP success.<br>P+ and P-  failed  -> retry.</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0821" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/21</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Predenature</td><td>95℃</td><td>3min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>98℃</td><td>20sec</td><td>25</td></tr>
 +
<tr><td>Annealing</td><td>70℃</td><td>15sec</td><td>25</td></tr>
 +
<tr><td>Extension</td><td>72℃</td><td>30sec</td><td>25</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>4℃</td><td>-</td><td>1</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0821" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/21</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>scfv 8/20 transformation</td><td>cscfv.f /0.75</td><td>cscfv.r /0.75</td><td>2x KAPA Hifi /12.5</td><td>11</td><td></td><td>25</td></tr>
 +
<tr><td>- (nega)</td><td>cscfv.f /0.3</td><td>cscfv.f /0.3</td><td>2x KAPA Hifi /5</td><td>4.4</td><td></td><td>10</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0821" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/21</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Colony</th><th>Primers</th></tr>
 +
<tr><td>8/20 transformation p+ [1]</td><td>VF+VR</td></tr>
 +
<tr><td>8/20 transformation p+ [2]</td><td>VF+VR</td></tr>
 +
<tr><td>8/20 transformation p+ [3]</td><td>VF+VR</td></tr>
 +
<tr><td>8/20 transformation p- [1]</td><td>VF+VR</td></tr>
 +
<tr><td>8/20 transformation p- [2]</td><td>VF+VR</td></tr>
 +
<tr><td>8/20 transformation p- [3]</td><td>VF+VR</td></tr>
 +
<tr><td>8/18 parts p+</td><td>VF+VR</td></tr>
 +
<tr><td>8/18 parts p-</td><td>VF+VR</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0821" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/21</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>pre</td><td>94℃</td><td>2min</td><td>1</td></tr>
 +
<tr><td>de</td><td>94℃</td><td>30sec</td><td>30</td></tr>
 +
<tr><td>an</td><td>55℃</td><td>30sec</td><td>30</td></tr>
 +
<tr><td>ex</td><td>68℃</td><td>1min</td><td>30</td></tr>
 +
<tr><td>F.elong</td><td>4℃</td><td>7min</td><td>1</td></tr>
 +
<tr><td>F.hold</td><td>4℃</td><td>-</td><td>1</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kaneko -->
 +
<div class="keln_container">
 +
<a name="0908" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/8</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td></td><td>[1] 8/15 miniprep 1c3 RFP /14</td><td>0.5</td><td></td><td>0.5</td><td>-</td><td>10xH /3</td><td></td><td>12</td><td>30</td></tr>
 +
<tr><td></td><td>[2] 8/24 miniprep 3c5 RFP /8.5</td><td>0.5</td><td></td><td>-</td><td>0.5</td><td>10xH /3</td><td></td><td>17.5</td><td>30</td></tr>
 +
<tr><td></td><td>[3] 8/24 miniprep 3c5 RFP /8.5</td><td>0.5</td><td></td><td>0.5</td><td>-</td><td>10xH /3</td><td></td><td>17.5</td><td>30</td></tr>
 +
<tr><td></td><td>[4] 8/24 miniprep BBa_J61002 p- /24</td><td>0.5</td><td></td><td>-</td><td>0.5</td><td>10xH /3</td><td></td><td>2</td><td>30</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/4/4c/T--Kyoto--9_8_No50_Restriction_Enzyme_Digestion_Kaneko_05_20_05_48_28.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto,Iguchi -->
 +
<div class="keln_container">
 +
<a name="0909" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/9</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Products</th></tr>
 +
<tr><td>1</td><td>100bp ladder 9&microL</td></tr>
 +
<tr><td>2</td><td>8/24 miniprep p- EPcut 30&microL</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto,Iguchi -->
 +
<div class="keln_container">
 +
<a name="0909" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/9</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Sukegawa, Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Products</th></tr>
 +
<tr><td>1</td><td>1kbp ladder 9&microL</td></tr>
 +
<tr><td>2</td><td>8/15 miniprep 1C3RFP EScut 30&microL</td></tr>
 +
<tr><td>3</td><td>8/24 miniprep 3C5RFP EPcut 30&microL</td></tr>
 +
<tr><td>4</td><td>8/24 miniprep 3C5RFP EScut 30&microL</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto -->
 +
<div class="keln_container">
 +
<a name="0914" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/14</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>1</td><td>95</td><td>3min</td><td>１</td></tr>
 +
<tr><td>2</td><td>98</td><td>70sec</td><td>25</td></tr>
 +
<tr><td>3</td><td>68</td><td>15sec</td><td>25</td></tr>
 +
<tr><td>4</td><td>72</td><td>30sec</td><td>25</td></tr>
 +
<tr><td>5</td><td>72</td><td>5min</td><td>1</td></tr>
 +
<tr><td>6</td><td>10</td><td>∞</td><td>1</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto -->
 +
<div class="keln_container">
 +
<a name="0914" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/14</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>8/24 miniprep INP</td><td>INPfw/0.75</td><td>INPRv/0.75</td><td>2×KAPAHiFi</td><td>11</td><td></td><td>25</td></tr>
 +
<tr><td>8/24 miniprep INP</td><td>INPfw/0.75</td><td>INPHisRv/0.75</td><td>2×KAPAHiFi</td><td>11</td><td></td><td>25</td></tr>
 +
<tr><td>BAN</td><td>BANFw/0/75</td><td>BANRv/0.75</td><td>2×KAPAHiFi</td><td>11</td><td></td><td>25</td></tr>
 +
<tr><td>BAN</td><td>BANFw/0/75</td><td>BANHisRv/0.75</td><td>2×KAPAHiFi</td><td>11</td><td></td><td>25</td></tr>
 +
<tr><td>BAN </td><td>BANFw/0/75</td><td>RBSRv/0.75</td><td>2×KAPAHiFi</td><td>11</td><td></td><td>25</td></tr>
 +
<tr><td>scFv fromMk</td><td>scFvFw/0/75</td><td>scFvRv/0/75</td><td>2×KAPAHiFi</td><td>11</td><td></td><td>25</td></tr>
 +
<tr><td>scFv fromMk</td><td>scFvHisFw/0.75</td><td></td><td>2×KAPAHiFi</td><td>11</td><td></td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto,Iguchi -->
 +
<div class="keln_container">
 +
<a name="0914" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/14</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Kaneko</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Products</th></tr>
 +
<tr><td>2</td><td>ladder/6</td></tr>
 +
<tr><td>4</td><td>9/14 PCR1/10</td></tr>
 +
<tr><td>6</td><td>9/14 PCR2/10</td></tr>
 +
<tr><td>8</td><td>9/14 PCR3/10</td></tr>
 +
<tr><td>10</td><td>9/14 PCR4/10</td></tr>
 +
<tr><td>12</td><td>9/14PCR5/10</td></tr>
 +
<tr><td>14</td><td>9/14 PCR6/10</td></tr>
 +
<tr><td>16</td><td>9/14 PCR7/10</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/4/4b/T--Kyoto--9_14_No53_Electrophoresis_Yamamoto_08_23_11_27_01.1.JPG"><img src="https://static.igem.org/mediawiki/2016/7/77/T--Kyoto--9_14_No53_Electrophoresis_Yamamoto_08_23_11_27_01.2.JPG">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto -->
 +
<div class="keln_container">
 +
<a name="0914" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/14</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3sBAN/0.15+6scFvfromMK/0.15</td><td>BANFw/0/75</td><td>scFvRv/0.75</td><td>2×KAPAHiFi/12.5</td><td></td><td></td><td>25</td></tr>
 +
<tr><td>4sBAN/0.15+7scFvfromMK/0.15</td><td>BANFw/0/75</td><td>scFvRv/0.75</td><td>2×KAPAHiFi/12.5</td><td></td><td></td><td>25</td></tr>
 +
<tr><td>1INP8/24miniprep/0.15+5sBAN0.15+scFvfromMK/0.15</td><td>BANFw/0/75</td><td>scFvRv/0.75</td><td>2×KAPAHiFi/12.5</td><td></td><td></td><td>25</td></tr>
 +
<tr><td>2INP8/24miniprep/0.15</td><td>BANFw/0/75</td><td>scFvRv/0.75</td><td>2×KAPAHiFi/12.5</td><td></td><td></td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto,Iguchi -->
 +
<div class="keln_container">
 +
<a name="0914" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/15</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Li, Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3sBAN+6scFvfromMKPCRproduct</td><td>34</td><td>1</td><td></td><td>1</td><td></td><td>4</td><td></td><td></td><td>40</td></tr>
 +
<tr><td>4sBAN+7scFvfromMK PCRproduct</td><td>34</td><td>1</td><td></td><td>1</td><td></td><td>4</td><td></td><td></td><td>40</td></tr>
 +
<tr><td>1INP8/24miniprep+5sBAN+6scFvfromMKPCRproduct</td><td>16</td><td>1</td><td></td><td>1</td><td></td><td>2</td><td></td><td></td><td>20</td></tr>
 +
<tr><td>2INP8/24miniprep+5sBAN+7scFvfromMk PCRproduct</td><td></td><td>1</td><td></td><td>1</td><td></td><td>4</td><td></td><td></td><td>40</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto -->
 +
<div class="keln_container">
 +
<a name="0915" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/15</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Li, Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td></td></tr>
 +
<tr><td>2</td><td>3sBAN+6scFvfromMkPCR産物EScut</td><td>20</td></tr>
 +
<tr><td>3</td><td>3sBAN+6scFvfromMkPCR産物EScut</td><td>20</td></tr>
 +
<tr><td>4</td><td>4sBAN+7scFvfromMkPCR産物EScut</td><td>20</td></tr>
 +
<tr><td>5</td><td>4sBAN+7scFvfromMkPCR産物EScut</td><td>20</td></tr>
 +
<tr><td>6</td><td>1INP8/24miniprep+5sBAN+7scFvfromMkPCR産物EScut</td><td>20</td></tr>
 +
<tr><td>7</td><td>2INP8/24miniprep+5sBAN+7scFvfromMkPCR産物EScut</td><td>20</td></tr>
 +
<tr><td>8</td><td>2INP8/24miniprep+5sBAN+7scFvfromMkPCR産物EScut</td><td>20</td></tr>
 +
<tr><td>9</td><td>RFP3C5PCR産物</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/a/a2/T--Kyoto--9_15_No56_Gel_Extraction_Yamamoto_08_23_11_46_18%2C1.jpg"><img src="https://static.igem.org/mediawiki/2016/5/52/T--Kyoto--9_15_No56_Gel_Extraction_Yamamoto_08_23_11_46_18%2C2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto -->
 +
<div class="keln_container">
 +
<a name="0916" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/16</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (2Column)</h4></span>
 +
<span class="keln_researcher">Uchino, Michimori</span>
 +
<table class="keln_table">
 +
<tr><th>Name I</th><th>Name II</th></tr>
 +
<tr><td>9/15 transformation BANscFv 3C5 colony1~16</td><td>VF+VR</td></tr>
 +
<tr><td>9/15 transformation INPscFv 3C5 colony1~16</td><td>VF+VR</td></tr>
 +
<tr><td>9/15 transformation BANHisscFv 3C5 colony1~16</td><td>VF+VR</td></tr>
 +
<tr><td>9/15 transformation INPHisscFv 3C5 colony1~16</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto -->
 +
<div class="keln_container">
 +
<a name="0916" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/16</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Uchino, Michimori</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>1</td><td>94</td><td>2</td><td>1</td></tr>
 +
<tr><td>2</td><td>94</td><td>0.5</td><td>25</td></tr>
 +
<tr><td>3</td><td>55</td><td>0.5</td><td>25</td></tr>
 +
<tr><td>4</td><td>68</td><td>1</td><td>25</td></tr>
 +
<tr><td>5</td><td>4</td><td>7</td><td>1</td></tr>
 +
<tr><td>6</td><td>4</td><td>∞</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto -->
 +
<div class="keln_container">
 +
<a name="0916" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/16</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Li, Kaneko, Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td>5</td></tr>
 +
<tr><td>2</td><td>BANscFv3C5 colony 9</td><td>10</td></tr>
 +
<tr><td>3</td><td>BANscFv3C5 colony 10</td><td>10</td></tr>
 +
<tr><td>4</td><td>BANscFv3C5 colony 11</td><td>10</td></tr>
 +
<tr><td>5</td><td>BANscFv3C5 colony 12</td><td>10</td></tr>
 +
<tr><td>6</td><td>BANscFv3C5 colony 13</td><td>10</td></tr>
 +
<tr><td>7</td><td>BANscFv3C5 colony 14</td><td>10</td></tr>
 +
<tr><td>8</td><td>BANscFv3C5 colony 15</td><td>10</td></tr>
 +
<tr><td>9</td><td>BANscFv3C5 colony 16</td><td>10</td></tr>
 +
<tr><td>10</td><td>BANHisscFv3C5 colony 9</td><td>10</td></tr>
 +
<tr><td>11</td><td>BANHisscFv3C5 colony 10</td><td>10</td></tr>
 +
<tr><td>12</td><td>BANHisscFv3C5 colony 11</td><td>10</td></tr>
 +
<tr><td>13</td><td>BANHisscFv3C5 colony 12</td><td>10</td></tr>
 +
<tr><td>14</td><td>BANHisscFv3C5 colony 13</td><td>10</td></tr>
 +
<tr><td>15</td><td>BANHisscFv3C5 colony 14</td><td>10</td></tr>
 +
<tr><td>16</td><td>BANHisscFv3C5 colony 15</td><td>10</td></tr>
 +
<tr><td>17</td><td>BANHisscFv3C5 colony 16</td><td>10</td></tr>
 +
<tr><td>18</td><td>BANscFv3C5 colony 1</td><td>10</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/d/d3/T--Kyoto--9_16_No59_Gel_Extraction_Yamamoto_08_23_11_57_19.1.JPG"><img src="https://static.igem.org/mediawiki/2016/3/34/T--Kyoto--9_16_No59_Gel_Extraction_Yamamoto_08_23_11_57_19.2.JPG"><img src="https://static.igem.org/mediawiki/2016/a/aa/T--Kyoto--9_16_No59_Gel_Extraction_Yamamoto_08_23_11_57_19.3.JPG"><img src="https://static.igem.org/mediawiki/2016/6/67/T--Kyoto--9_16_No59_Gel_Extraction_Yamamoto_08_23_11_57_19.4.JPG">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto -->
 +
<div class="keln_container">
 +
<a name="0916" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/16</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Li, Kaneko, Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td>5</td></tr>
 +
<tr><td>2</td><td>INPscFv3C5 colony 9</td><td>10</td></tr>
 +
<tr><td>3</td><td>INPscFv3C5 colony 10</td><td>10</td></tr>
 +
<tr><td>4</td><td>INPscFv3C5 colony 11</td><td>10</td></tr>
 +
<tr><td>5</td><td>INPscFv3C5 colony 12</td><td>10</td></tr>
 +
<tr><td>6</td><td>INPscFv3C5 colony 13</td><td>10</td></tr>
 +
<tr><td>7</td><td>INPscFv3C5 colony 14</td><td>10</td></tr>
 +
<tr><td>8</td><td>INPscFv3C5 colony 15</td><td>10</td></tr>
 +
<tr><td>9</td><td>INPscFv3C5 colony 16</td><td>10</td></tr>
 +
<tr><td>10</td><td>INPHisscFv3C5 colony 9</td><td>10</td></tr>
 +
<tr><td>11</td><td>INPHisscFv3C5 colony 10</td><td>10</td></tr>
 +
<tr><td>12</td><td>INPHisscFv3C5 colony 11</td><td>10</td></tr>
 +
<tr><td>13</td><td>INPHisscFv3C5 colony 12</td><td>10</td></tr>
 +
<tr><td>14</td><td>INPHisscFv3C5 colony 13</td><td>10</td></tr>
 +
<tr><td>15</td><td>INPHisscFv3C5 colony 14</td><td>10</td></tr>
 +
<tr><td>16</td><td>INPHisscFv3C5 colony 15</td><td>10</td></tr>
 +
<tr><td>17</td><td>INPHisscFv3C5 colony 16</td><td>10</td></tr>
 +
<tr><td>18</td><td>BANscFv3C5 colony 1</td><td>10</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/3/3d/T--Kyoto--9_16_No60_Gel_Extraction_Yamamoto_08_23_11_59_27%2C1.jpg"><img src="https://static.igem.org/mediawiki/2016/8/88/T--Kyoto--9_16_No60_Gel_Extraction_Yamamoto_08_23_11_59_27%2C2.jpg"><img src=""><img src="https://static.igem.org/mediawiki/2016/4/42/T--Kyoto--9_16_No60_Gel_Extraction_Yamamoto_08_23_11_59_27%2C3.jpg"><img src="https://static.igem.org/mediawiki/2016/6/69/T--Kyoto--9_16_No60_Gel_Extraction_Yamamoto_08_23_11_59_27%2C4.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto,Iguchi -->
 +
<div class="keln_container">
 +
<a name="0916" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/17</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Michimori, Kaneko</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Products</th></tr>
 +
<tr><td>1</td><td>buffer/5</td></tr>
 +
<tr><td>2</td><td>1kb ladder/5</td></tr>
 +
<tr><td>3</td><td>BAN 9 P/4.5</td></tr>
 +
<tr><td>4</td><td>BAN 9 N/4.5</td></tr>
 +
<tr><td>5</td><td>BAN14 P/4.5</td></tr>
 +
<tr><td>6</td><td>BAN14 N/4.5</td></tr>
 +
<tr><td>7</td><td>BANHis14 P/4.5</td></tr>
 +
<tr><td>8</td><td>BANHis14/4.5</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/9/98/T--Kyoto--9_17_No62_Electrophoresis_Yamamoto_08_23_12_42_58.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Iguchi -->
 +
<div class="keln_container">
 +
<a name="0917" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/17</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Kaneko</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kbp ladder</td><td></td></tr>
 +
<tr><td>2</td><td>none</td><td></td></tr>
 +
<tr><td>3</td><td>BANHis16 p</td><td></td></tr>
 +
<tr><td>4</td><td>BANHis16 n</td><td></td></tr>
 +
<tr><td>5</td><td>INP 12 p</td><td></td></tr>
 +
<tr><td>6</td><td>INP 14 p</td><td></td></tr>
 +
<tr><td>7</td><td>INP 12 n</td><td></td></tr>
 +
<tr><td>8</td><td></td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/9/9b/T--Kyoto--9_17_No63_Electrophoresis_Iguchi_10_17_06_39_35.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto,Iguchi -->
 +
<div class="keln_container">
 +
<a name="0917" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/17</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Kaneko</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Products</th></tr>
 +
<tr><td>1</td><td>-</td></tr>
 +
<tr><td>2</td><td>-</td></tr>
 +
<tr><td>3</td><td>1kbladder/5</td></tr>
 +
<tr><td>4</td><td>INPHis9 P/4.5</td></tr>
 +
<tr><td>5</td><td>INPHis9N/4.5</td></tr>
 +
<tr><td>6</td><td>INPHis12P/4.5</td></tr>
 +
<tr><td>7</td><td>INPHis12N/4.5</td></tr>
 +
<tr><td>8</td><td>INP14 N/4.5</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/b/b8/T--Kyoto--9_17_No64_Electrophoresis_Yamamoto_08_23_01_05_58.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto -->
 +
<div class="keln_container">
 +
<a name="0917" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/17</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Uchino, Michimori</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>BAN9 </td><td>2</td><td>0.5</td><td></td><td></td><td>0.5</td><td>10×H/1</td><td></td><td>6</td><td>10</td></tr>
 +
<tr><td>BAN9 </td><td>1</td><td>0</td><td></td><td></td><td>0</td><td>10×H/1</td><td></td><td>8</td><td>10</td></tr>
 +
<tr><td>BAN14 </td><td>2</td><td>0.5</td><td></td><td></td><td>0.5</td><td>10×H/1</td><td></td><td>6</td><td>10</td></tr>
 +
<tr><td>BAN14 </td><td>1</td><td>0</td><td></td><td></td><td>0</td><td>10×H/1</td><td></td><td>8</td><td>10</td></tr>
 +
<tr><td>BANHis14 </td><td>2</td><td>0.5</td><td></td><td></td><td>0.5</td><td>10×H/1</td><td></td><td>6</td><td>10</td></tr>
 +
<tr><td>BANHis 14 </td><td>1</td><td>0</td><td></td><td></td><td>0</td><td>10×H/1</td><td></td><td>8</td><td>10</td></tr>
 +
<tr><td>BANHis 16 </td><td>2</td><td>0.5</td><td></td><td></td><td>0.5</td><td>10×H/1</td><td></td><td>6</td><td>10</td></tr>
 +
<tr><td>BANHis16 </td><td>1</td><td>0</td><td></td><td></td><td>0</td><td>10×H/1</td><td></td><td>8</td><td>10</td></tr>
 +
<tr><td>INP12 </td><td>2</td><td>0.5</td><td></td><td></td><td>0.5</td><td>10×H/1</td><td></td><td>6</td><td>10</td></tr>
 +
<tr><td>INP12 </td><td>1</td><td>0</td><td></td><td></td><td>0</td><td>10×H/1</td><td></td><td>8</td><td>10</td></tr>
 +
<tr><td>INP14 </td><td>2</td><td>0.5</td><td></td><td></td><td>0.5</td><td>10×H/1</td><td></td><td>6</td><td>10</td></tr>
 +
<tr><td>INP 14 </td><td>1</td><td>0</td><td></td><td></td><td>0</td><td>10×H/1</td><td></td><td>8</td><td>10</td></tr>
 +
<tr><td>INP 9 </td><td>2</td><td>0.5</td><td></td><td></td><td>0.5</td><td>10×H/1</td><td></td><td>6</td><td>10</td></tr>
 +
<tr><td>INP9 </td><td>1</td><td>0</td><td></td><td></td><td>0</td><td>10×H/1</td><td></td><td>8</td><td>10</td></tr>
 +
<tr><td>INPHis12 </td><td>2</td><td>0.5</td><td></td><td></td><td>0.5</td><td>10×H/1</td><td></td><td>6</td><td>10</td></tr>
 +
<tr><td>INPHis12 </td><td>1</td><td>0</td><td></td><td></td><td>0</td><td>10×H/1</td><td></td><td>8</td><td>10</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/8/8a/T--Kyoto--9_17_No65_Restriction_Enzyme_Digestion_Yamamoto_08_23_01_13_52.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto -->
 +
<div class="keln_container">
 +
<a name="0917" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/17</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>9/16 liquid culture BANscFv12</td><td></td><td></td><td></td></tr>
 +
<tr><td>9/16 liquid culture BANscFv14</td><td></td><td></td><td></td></tr>
 +
<tr><td>9/16 liquid culture BANHisscFv12</td><td></td><td></td><td></td></tr>
 +
<tr><td>9/16 liquid culture BANHisscFv14</td><td></td><td></td><td></td></tr>
 +
<tr><td>9/16 liquid culture INPHisscFv12</td><td></td><td></td><td></td></tr>
 +
<tr><td>9/16 liquid culture INPHisscFv14</td><td></td><td></td><td></td></tr>
 +
<tr><td>9/16 liquid culture INPHisscFv10</td><td></td><td></td><td></td></tr>
 +
<tr><td>9/16 liquid culture BANscFv</td><td></td><td></td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0204" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/4</h3></span>
 +
<span class="keln_exp"><h4>PartsAwaking</h4></span>
 +
<span class="keln_researcher">Kaneko, Wan</span>
 +
<table class="keln_table">
 +
<tr><th>Description</th><th>PartName</th><th>Backbone</th><th>Well</th><th>PlateYear</th><th>PlateNumber</th><th>MilliQ(&microl)</th></tr>
 +
<tr><td>RFP</td><td>E1010</td><td>pSB1C3</td><td>11N</td><td>2015</td><td>3</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0204" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/4</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Uchino, Iguchi, Notsu, Wan</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>E1010</td><td>2</td><td>DH5α</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0205" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/5</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Uchino, Egashira, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/4 transformation E1010 colony1</td><td>VF 6.25&microl      VR 6.25&microl</td><td>12.5</td><td>2×Quick Taq HS DyeMix 25&microl</td><td>50</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0205" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/5</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Uchino, Egashira, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Predenature</td><td>94℃</td><td>2min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>94℃</td><td>30sec</td><td>25</td></tr>
 +
<tr><td>Annealing</td><td>51℃</td><td>30sec</td><td>25</td></tr>
 +
<tr><td>Extension</td><td>68℃</td><td>90sec</td><td>25</td></tr>
 +
<tr><td>Final Elongation</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
<tr><td>Final Holding</td><td>4℃</td><td>-</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto -->
 +
<div class="keln_container">
 +
<a name="0205" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/5</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>9/17 miniprep BAN9</td><td>2</td><td>DH5α/20&microL</td><td></td></tr>
 +
<tr><td>9/17　miniprep BANHis14</td><td>2</td><td>DH5α/20&microL</td><td></td></tr>
 +
<tr><td>9/17miniprep INPHis12</td><td>2</td><td>DH5α/20&microL</td><td></td></tr>
 +
<tr><td>9/17miniprep INPHis9</td><td>2</td><td></td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0205" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/5</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Uchino, Wan, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>2/4 transformation E1010 colony1</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 
<!-- Table Generated by KELN Author: Notsu -->
 
<!-- Table Generated by KELN Author: Notsu -->
 
<div class="keln_container">
 
<div class="keln_container">
Line 189: Line 1,491:
 
</div>
 
</div>
 
<!------------ Table END ------------>
 
<!------------ Table END ------------>
−
 
+
<!-- Table Generated by KELN Author: Notsu -->
−
<!-- Table Generated by KELN Author: Sukegawa -->
+
−
 
+
 
<div class="keln_container">
 
<div class="keln_container">
−
 
+
<a name="0206" class = "kyoto-jump"></a>
−
<a name="0101" class = "kyoto-jump"></a>
+
<span class="keln_date"><h3>2/6</h3></span>
−
 
+
<span class="keln_exp"><h4>Miniprep</h4></span>
−
<span class="keln_date"><h3>1/1</h3></span>
+
<span class="keln_researcher">Notsu, Kaneko, Egashira, Wan, Yamamoto</span>
−
 
+
−
<span class="keln_exp"><h4>PCR (Target)</h4></span>
+
−
 
+
−
<span class="keln_researcher">Sukegawa</span>
+
−
 
+
 
<table class="keln_table">
 
<table class="keln_table">
−
 
+
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
−
<tr><th>Templates/(μl)</th><th>Primer1/(μl)</th><th>Primer2/(μl)</th><th>Buffer/(μl)</th><th>MilliQ/(μl)</th><th>Any Other/(μl)</th><th>Total/(μl)</th></tr>
+
<tr><td>2/5 liquid culture E1010(RFP)  colony1</td><td>48.0</td><td>1.93</td><td></td></tr>
−
 
+
−
<tr><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
+
−
 
+
 
</table>
 
</table>
−
 
 
</div>
 
</div>
−
 
 
<!------------ Table END ------------>
 
<!------------ Table END ------------>
−
 
+
<!-- Table Generated by KELN Author: Notsu -->
−
<!-- Table Generated by KELN Author: Sukegawa -->
+
−
 
+
 
<div class="keln_container">
 
<div class="keln_container">
−
 
+
<a name="0206" class = "kyoto-jump"></a>
−
<a name="0101" class = "kyoto-jump"></a>
+
<span class="keln_date"><h3>2/6</h3></span>
−
 
+
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
−
<span class="keln_date"><h3>1/1</h3></span>
+
<span class="keln_researcher">Kaneko, Egashira, Wan, Yamamoto</span>
−
 
+
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/6 miniprep E1010 colony1</td><td>34</td><td>0</td><td>1</td><td>0</td><td>1</td><td>1×M /4</td><td>0</td><td>0</td><td>40</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0207" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/7</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Kaneko, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder </td><td>10</td></tr>
 +
<tr><td>2</td><td>E1010 colony1 2/6 XPcut</td><td>40</td></tr>
 +
<tr><td>3</td><td>other sample</td><td>-</td></tr>
 +
<tr><td>4</td><td>other sample</td><td>-</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/8/84/T--Kyoto--2_7_No78_Gel_Extraction_Notsu_10_01_04_37_25.1.JPG"><img src="https://static.igem.org/mediawiki/2016/6/68/T--Kyoto--2_7_No78_Gel_Extraction_Notsu_10_01_04_37_25.2.JPG">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0207" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/7</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Kaneko, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>E1010 colony1 2/6 XPcut</td><td>8.2</td><td>1.03</td><td>0.49</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/7/71/T--Kyoto--2_7_No79_Gel_Extraction%28Measurement%29_Notsu_10_01_04_40_48.jpg">
 +
<img src="https://static.igem.org/mediawiki/2016/7/71/T--Kyoto--2_7_No79_Gel_Extraction%28Measurement%29_Notsu_10_01_04_40_48.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0207" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/7</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>2/5 transformation BAN9 colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>2/5 transformation BAN-HIS14 colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>2/5 transformation INP-HIS12 colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>2/5 transformation BAN9 colony1</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0208" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/8</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Uchino, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>2/4 transformation E1010 colony2 1</td><td>LB(CP)</td></tr>
 +
<tr><td>2/4 transformation E1010 colony2 2</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0208" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/8</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Uchino, Egashira, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/7 Liquid culture BAN9</td><td>123.6</td><td>1.87</td><td>2.07</td></tr>
 +
<tr><td>2/7 Liquid culture BAN9</td><td>248.8</td><td>1.83</td><td>1.84</td></tr>
 +
<tr><td>2/7 Liquid culture BAN-HIS14</td><td>268.9</td><td>1.86</td><td>2.15</td></tr>
 +
<tr><td>2/7 Liquid culture BAN-HIS14</td><td>127.9</td><td>1.85</td><td>2.08</td></tr>
 +
<tr><td>2/7 Liquid culture INP-HIS9</td><td>354.3</td><td>1.86</td><td>2.24</td></tr>
 +
<tr><td>2/7 Liquid culture INP-HIS9</td><td>289.4</td><td>1.85</td><td>1.92</td></tr>
 +
<tr><td>2/7 Liquid culture INP-HIS12</td><td>172.3</td><td>1.87</td><td>2.07</td></tr>
 +
<tr><td>2/7 Liquid culture INP-HIS12</td><td>286.5</td><td>1.86</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0208" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/8</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino, Egashira, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /5</td><td></td></tr>
 +
<tr><td>2</td><td>2/8 miniprep BAN9 /5</td><td></td></tr>
 +
<tr><td>3</td><td>2/8 miniprep BAN9 /5</td><td></td></tr>
 +
<tr><td>4</td><td>2/8 miniprep BAN-HIS14 /5</td><td></td></tr>
 +
<tr><td>5</td><td>2/8 miniprep BAN-HIS14 /5</td><td></td></tr>
 +
<tr><td>6</td><td>2/8 miniprep INP-HIS9 /5</td><td></td></tr>
 +
<tr><td>7</td><td>2/8 miniprep INP-HIS9 /5</td><td></td></tr>
 +
<tr><td>8</td><td>2/8 miniprep INP-HIS14 /5</td><td></td></tr>
 +
<tr><td>9</td><td>2/8 miniprep INP-HIS9 /5</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/0/0b/T--Kyoto--2_8_No85_Electrophoresis_Notsu_09_29_01_38_11.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0209" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/9</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Matsumoto, Notsu, Egashira, Yamamoto, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/8 liquid culture E1010 colony2 1and 2</td><td>58.5</td><td>1.89</td><td>2.16</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0209" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/9</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Uchino, Egashira, Yamamoto, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/8 liquid culture E1010 colony2 </td><td>30</td><td>0</td><td>1</td><td>0</td><td>1</td><td>10×M /4</td><td>2</td><td>2</td><td>40</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0209" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/9</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Uchino, Yamamoto, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/8 liquid culture E1010 colony2 </td><td>5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>10×M /2</td><td>1</td><td>12</td><td>20</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Takimoto -->
 +
<div class="keln_container">
 +
<a name="0210" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/10</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino, Yamamoto, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /5</td><td></td></tr>
 +
<tr><td>2</td><td>2/9 Negative Control E1010 colony2 /10</td><td>2776</td></tr>
 +
<tr><td>3</td><td>2/9 Digestion E1010 colony2 p+x /40</td><td>732</td></tr>
 +
<tr><td>4</td><td>2/9 Negative Control E0040 colony2 /20</td><td>2799</td></tr>
 +
<tr><td>5</td><td>2/9 Digestion E0040 colony2 p+x /40</td><td>746</td></tr>
 +
<tr><td>6</td><td>2/9 Negative Control K592009 colony2 /20</td><td>2739</td></tr>
 +
<tr><td>7</td><td>2/9 Digestion K592009 colony2 p+x /40</td><td>695</td></tr>
 +
<tr><td>8</td><td>2/9 Negative Control E0022 colony2 /20</td><td>2832</td></tr>
 +
<tr><td>9</td><td>2/9 Digestion E0022 colony2 p+x /40</td><td></td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">Lane7流出</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Takimoto -->
 +
<div class="keln_container">
 +
<a name="0210" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/10</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino, Yamamoto, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /5</td><td></td></tr>
 +
<tr><td>3</td><td>2/9 NC J23105 colony2 /20</td><td>2105</td></tr>
 +
<tr><td>4</td><td>2/9 Digestion J23105 colony2 E+S /40</td><td>58</td></tr>
 +
<tr><td>5</td><td>2/9 NC J23100 colony2 /20</td><td>2105</td></tr>
 +
<tr><td>6</td><td>2/9 Digestion J23100 colony2 E+S /40</td><td>58</td></tr>
 +
<tr><td>7</td><td>2/9 NC B0034 colony3 /20</td><td>2091</td></tr>
 +
<tr><td>8</td><td>2/9 Digestion B0034 colony3 P+S /40</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/4/48/T--Kyoto--2_10_No91_Electrophoresis_Takimoto_09_14_05_48_30.1.JPG"><img src="https://static.igem.org/mediawiki/2016/4/45/T--Kyoto--2_10_No91_Electrophoresis_Takimoto_09_14_05_48_30.2.JPG"><img src="https://static.igem.org/mediawiki/2016/7/73/T--Kyoto--2_10_No91_Electrophoresis_Takimoto_09_14_05_48_30.3.JPG"><img src="https://static.igem.org/mediawiki/2016/b/b0/T--Kyoto--2_10_No91_Electrophoresis_Takimoto_09_14_05_48_30.4.JPG">
 +
</div>
 +
<p class="keln_note">2には何も入れてない</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Takimoto -->
 +
<div class="keln_container">
 +
<a name="0210" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/10</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Michimori, Egashira, Yamamoto, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>E1010</td><td>9.4</td><td>2.13</td><td>0.61</td></tr>
 +
<tr><td>K592009</td><td>5.2</td><td>1.74</td><td>0.29</td></tr>
 +
<tr><td>E0040</td><td>15.0</td><td>1.94</td><td>0.38</td></tr>
 +
<tr><td>E0022</td><td>8.5</td><td>1.04</td><td>0.86</td></tr>
 +
<tr><td>B0034</td><td>21.2</td><td>1.84</td><td>1.06</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">E1010　理想値18.5ng/&microLぐらい...<br>(制限酵素処理100%、ゲル抽出80%として)</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0212" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/12</h3></span>
 +
<span class="keln_exp"><h4>PartsAwaking</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Description</th><th>PartName</th><th>Backbone</th><th>Well</th><th>PlateYear</th><th>PlateNumber</th><th>MilliQ(&microl)</th></tr>
 +
<tr><td>sfGFP-CBDcex</td><td>K1321342</td><td>pSB1C3</td><td>18E</td><td>2015</td><td>5</td><td>10</td></tr>
 +
<tr><td>sfGFP-CBDclos</td><td>K1321341</td><td>pSB1C3</td><td>18C</td><td>2015</td><td>5</td><td>10</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0212" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/12</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>2/12 parts awakening sfGFP-CBDcex</td><td>1</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>2/12 parts awakening sfGFP-CBDclos</td><td>1</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0212" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/12</h3></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
−
 
+
<span class="keln_researcher">Uchino</span>
−
<span class="keln_researcher">Sukegawa</span>
+
−
 
+
 
<table class="keln_table">
 
<table class="keln_table">
−
 
+
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
−
<tr><th>Templates/(μl)</th><th>Primer1/(μl)</th><th>Primer2/(μl)</th><th>Buffer/(μl)</th><th>MilliQ/(μl)</th><th>Any Other/(μl)</th><th>Total/(μl)</th></tr>
+
<tr><td>2/8 miniprep BAN-His14 4 /0.1</td><td>VF /0.75</td><td>m.scFv.Rv /0.75</td><td>-</td><td>10.9</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
−
 
+
<tr><td>2/8 miniprep BAN-His14 4 /0.1</td><td>VR /0.75</td><td>m.scFv.Fw /0.75</td><td>-</td><td>10.9</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
−
<tr><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
+
</table>
−
 
+
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0212" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/12</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td>5</td></tr>
 +
<tr><td>2</td><td>2/12 PCR VF.m.scFv.Rv</td><td>25</td></tr>
 +
<tr><td>3</td><td>2/12 PCR VR.m.Fw</td><td>25</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/0/0a/T--Kyoto--2_12_No101_Electrophoresis_Notsu_10_01_07_30_03%2C1.jpg"> <img src="https://static.igem.org/mediawiki/2016/c/c8/T--Kyoto--2_12_No101_Electrophoresis_Notsu_10_01_07_30_03%2C2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0212" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/12</h3></span>
 +
<span class="keln_exp"><h4>PartsAwaking</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Description</th><th>PartName</th><th>Backbone</th><th>Well</th><th>PlateYear</th><th>PlateNumber</th><th>MilliQ(&microl)</th></tr>
 +
<tr><td>Promoter</td><td>J23114</td><td>J61002</td><td>19J</td><td>2015</td><td>4</td><td>10</td></tr>
 +
<tr><td>Promoter</td><td>J23108</td><td>pSB1C3</td><td>4C</td><td>2015</td><td>4</td><td>10</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0212" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/12</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Uchino, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>2/12 Parts awakening J23114</td><td>1</td><td>DH5α /20</td><td>LB(AMP)</td></tr>
 +
<tr><td>2/12 Parts awakening J23108</td><td>1</td><td>DH5α /20</td><td>LB(AMP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0212" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/12</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/12 PCR VF.m.scFv.Rv </td><td>18.8</td><td>1.95</td><td>1.52</td></tr>
 +
<tr><td>2/12 PCR VR.m.scFv.Fw </td><td>28.6</td><td>2.08</td><td>0.07</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0215" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/15</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/12 transformation sfGFP-CBDcex colony1</td><td>VF /0.75    VR /0.75</td><td>11</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
 +
<tr><td>2/12 transformation sfGFP-CBDcex colony2</td><td>VF /0.75    VR /0.75</td><td>11</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
 +
<tr><td>2/12 transformation sfGFP-CBDclos colony1</td><td>VF /0.75    VR /0.75</td><td>11</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
 +
<tr><td>2/12 transformation sfGFP-CBDclos colony2</td><td>VF /0.75    VR /0.75</td><td>11</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0215" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/15</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Kaneko</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /5</td><td></td></tr>
 +
<tr><td>2</td><td>2/12 transformation sfGFP-CBDcex colony1</td><td></td></tr>
 +
<tr><td>3</td><td>2/12 transformation sfGFP-CBDcex colony2</td><td></td></tr>
 +
<tr><td>4</td><td>2/12 transformation sfGFP-CBDclos colony1</td><td></td></tr>
 +
<tr><td>5</td><td>2/12 transformation sfGFP-CBDclos colony2</td><td></td></tr>
 
</table>
 
</table>
−
<img src="https://static.igem.org/mediawiki/2016/3/32/T--Kyoto--2_15_No3_Electrophoresis_Notsu_10_02_06_38_04.jpg">
 
 
<img src="https://static.igem.org/mediawiki/2016/3/32/T--Kyoto--2_15_No3_Electrophoresis_Notsu_10_02_06_38_04.jpg">
 
<img src="https://static.igem.org/mediawiki/2016/3/32/T--Kyoto--2_15_No3_Electrophoresis_Notsu_10_02_06_38_04.jpg">
 
</div>
 
</div>
−
 
 
<!------------ Table END ------------>
 
<!------------ Table END ------------>
−
 
+
<!-- Table Generated by KELN Author: Notsu -->
−
<!-- Table Generated by KELN Author: Sukegawa -->
+
−
 
+
 
<div class="keln_container">
 
<div class="keln_container">
−
 
+
<a name="0215" class = "kyoto-jump"></a>
−
<a name="0101" class = "kyoto-jump"></a>
+
<span class="keln_date"><h3>2/15</h3></span>
−
 
+
<span class="keln_exp"><h4>Liquid Culture</h4></span>
−
<span class="keln_date"><h3>1/1</h3></span>
+
<span class="keln_researcher">Wan, Yamamoto</span>
−
 
+
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>2/12 transformation sfGFP-CBDcex colony2</td><td>LB(CP)</td></tr>
 +
<tr><td>2/12 transformation sfGFP-CBDclos colony2</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0215" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/15</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/12 transformation J23114 colony1</td><td>VF 0.75          VR 0.75</td><td>11</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
 +
<tr><td>2/12 transformation J23114 colony2</td><td>VF 0.75          VR 0.75</td><td>11</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
 +
<tr><td>2/12 transformation J23108 colony1</td><td>VF 0.75          VR 0.75</td><td>11</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
 +
<tr><td>2/12 transformation J23108 colony2</td><td>VF 0.75          VR 0.75</td><td>11</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0215" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/15</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Predenature </td><td>95℃</td><td>3min</td><td>1</td></tr>
 +
<tr><td>Denature </td><td>98℃</td><td>20sec</td><td>35</td></tr>
 +
<tr><td>Annealing </td><td>65℃</td><td>15sec</td><td>35</td></tr>
 +
<tr><td>Extension </td><td>72℃</td><td>30sec</td><td>35</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0215" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/15</h3></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
−
 
+
<span class="keln_researcher">Uchino</span>
−
<span class="keln_researcher">Sukegawa</span>
+
−
 
+
 
<table class="keln_table">
 
<table class="keln_table">
−
 
+
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
−
<tr><th>Templates/(μl)</th><th>Primer1/(μl)</th><th>Primer2/(μl)</th><th>Buffer/(μl)</th><th>MilliQ/(μl)</th><th>Any Other/(μl)</th><th>Total/(μl)</th></tr>
+
<tr><td>2/12 Gel extraction PCR VF/m.scFv.Rv /0.53 + 2/12 Gel extraction PCR VR/m.scFv.Fw /0.35</td><td>VF /0.75</td><td>VR /0.75</td><td>-</td><td>10.12</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
−
 
+
<tr><td>2/12 Gel extraction PCR VF/m.scFv.Rv /0.53 + 2/12 Gel extraction PCR VR/m.scFv.Fw /0.35</td><td>VF /0.75</td><td>VR /0.75</td><td>-</td><td>10.12</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
−
<tr><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
+
−
 
+
 
</table>
 
</table>
−
 
 
</div>
 
</div>
−
 
 
<!------------ Table END ------------>
 
<!------------ Table END ------------>
−
 
+
<!-- Table Generated by KELN Author: Notsu -->
−
<!-- Table Generated by KELN Author: Sukegawa -->
+
−
 
+
 
<div class="keln_container">
 
<div class="keln_container">
−
 
+
<a name="0215" class = "kyoto-jump"></a>
−
<a name="0101" class = "kyoto-jump"></a>
+
<span class="keln_date"><h3>2/15</h3></span>
−
 
+
<span class="keln_exp"><h4>Electrophoresis</h4></span>
−
<span class="keln_date"><h3>1/1</h3></span>
+
<span class="keln_researcher">Kaneko</span>
−
 
+
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /5</td><td></td></tr>
 +
<tr><td>2</td><td>other sample</td><td>-</td></tr>
 +
<tr><td>3</td><td>other sample</td><td>-</td></tr>
 +
<tr><td>4</td><td>other sample</td><td>-</td></tr>
 +
<tr><td>5</td><td>other sample</td><td>-</td></tr>
 +
<tr><td>6</td><td>other sample</td><td>-</td></tr>
 +
<tr><td>7</td><td>other sample</td><td>-</td></tr>
 +
<tr><td>8</td><td>other sample</td><td>-</td></tr>
 +
<tr><td>9</td><td>other sample</td><td>-</td></tr>
 +
<tr><td>10</td><td>other sample</td><td>-</td></tr>
 +
<tr><td>11</td><td>other sample</td><td>-</td></tr>
 +
<tr><td>12</td><td>other sample</td><td>-</td></tr>
 +
<tr><td>13</td><td>other sample</td><td>-</td></tr>
 +
<tr><td>14</td><td>2/12 transformation J23114 colony1 /10</td><td>-</td></tr>
 +
<tr><td>15</td><td>2/12 transformation J23114 colony2 /10</td><td></td></tr>
 +
<tr><td>16</td><td>2/12 transformation J23108 colony1 /10</td><td></td></tr>
 +
<tr><td>17</td><td>2/12 transformation J23108 colony2 /10</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/6/6d/T--Kyoto--2_15_No112_Electrophoresis_Notsu_10_01_08_13_13.JPG">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0215" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/15</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1ka ladder</td><td>5</td></tr>
 +
<tr><td>2</td><td>2/15 OE-PCR VF/m.scFv.Rv+VR/m.scFv.Fw</td><td>25</td></tr>
 +
<tr><td>3</td><td>2/15 OE-PCR VF/m.scFv.Rv+VR/m.scFv.Fw</td><td>25</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/8/85/T--Kyoto--2_15_No114_Gel_Extraction_Notsu_10_01_08_21_26.1.JPG"><img src="https://static.igem.org/mediawiki/2016/3/3b/T--Kyoto--2_15_No114_Gel_Extraction_Notsu_10_01_08_21_26.2.JPG">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0215" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/15</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Wan, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>2/12 transformation J23114 colony2</td><td>LB(AMP)</td></tr>
 +
<tr><td>2/12 transformation J23108 colony2</td><td>LB(AMP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0216" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/16</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Wan, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/15 liquid culture sfGFP-CBDcex colony2</td><td>43.5</td><td>1.91</td><td>1.60</td></tr>
 +
<tr><td>2/15 liquid culture sfGFP-CBDclos colony2</td><td>74.0</td><td>1.86</td><td>1.82</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0216" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/16</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher"></span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Predenature</td><td>95℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>98℃</td><td>20sec</td><td>35</td></tr>
 +
<tr><td>Annealing</td><td>65℃</td><td>15sec</td><td>35</td></tr>
 +
<tr><td>Extension</td><td>72℃</td><td>30sec</td><td>35</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0216" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/16</h3></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
−
 
+
<span class="keln_researcher"></span>
−
<span class="keln_researcher">Sukegawa</span>
+
−
 
+
 
<table class="keln_table">
 
<table class="keln_table">
−
 
+
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
−
<tr><th>Templates/(μl)</th><th>Primer1/(μl)</th><th>Primer2/(μl)</th><th>Buffer/(μl)</th><th>MilliQ/(μl)</th><th>Any Other/(μl)</th><th>Total/(μl)</th></tr>
+
<tr><td>2/16 miniprep sfGFP-CBDcex colony2 /0.3</td><td>cex.Fw /0.75</td><td>VR /0.75</td><td>-</td><td>10.7</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
−
 
+
<tr><td>2/16 miniprep sfGFP-CBDclos colony2 /0.3</td><td>cex.Fw /0.75</td><td>VR /0.75</td><td>-</td><td>10.7</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
−
<tr><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
+
−
 
+
 
</table>
 
</table>
−
 
 
</div>
 
</div>
−
 
 
<!------------ Table END ------------>
 
<!------------ Table END ------------>
−
 
+
<!-- Table Generated by KELN Author: Notsu -->
−
<!-- Table Generated by KELN Author: Sukegawa -->
+
−
 
+
 
<div class="keln_container">
 
<div class="keln_container">
−
 
+
<a name="0216" class = "kyoto-jump"></a>
−
<a name="0101" class = "kyoto-jump"></a>
+
<span class="keln_date"><h3>2/16</h3></span>
−
 
+
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
−
<span class="keln_date"><h3>1/1</h3></span>
+
<span class="keln_researcher">Yamamoto</span>
−
 
+
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/15 OE-PCR VF/m.scFv.Rv+VR/m.scFv.Fw </td><td>48.6</td><td>1.83</td><td>0.19</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0216" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/16</h3></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
−
 
+
<span class="keln_researcher">Yamamoto, Yamada</span>
−
<span class="keln_researcher">Sukegawa</span>
+
−
 
+
 
<table class="keln_table">
 
<table class="keln_table">
−
 
+
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
−
<tr><th>Templates/(μl)</th><th>Primer1/(μl)</th><th>Primer2/(μl)</th><th>Buffer/(μl)</th><th>MilliQ/(μl)</th><th>Any Other/(μl)</th><th>Total/(μl)</th></tr>
+
<tr><td>2/15 Gel Extraction OE-PCR VF/m.scFv.Rv+VR/m.scFv.Fw /0.3</td><td>VF /0.75</td><td>VR /0.75</td><td>-</td><td>10.7</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
−
 
+
<tr><td>2/15 Gel Extraction OE-PCR VF/m.scFv.Rv+VR/m.scFv.Fw /0.3</td><td>VF /0.75</td><td>VR /0.75</td><td>-</td><td>10.7</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
−
<tr><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
+
−
 
+
 
</table>
 
</table>
−
 
 
</div>
 
</div>
−
 
 
<!------------ Table END ------------>
 
<!------------ Table END ------------>
−
 
+
<!-- Table Generated by KELN Author: Notsu -->
−
<!-- Table Generated by KELN Author: Sukegawa -->
+
−
 
+
 
<div class="keln_container">
 
<div class="keln_container">
−
 
+
<a name="0216" class = "kyoto-jump"></a>
−
<a name="0101" class = "kyoto-jump"></a>
+
<span class="keln_date"><h3>2/16</h3></span>
−
 
+
<span class="keln_exp"><h4>Miniprep</h4></span>
−
<span class="keln_date"><h3>1/1</h3></span>
+
<span class="keln_researcher">Uchino, Yamada</span>
−
 
+
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/15 liquid culture J23114</td><td>178.8</td><td>1.91</td><td>2.20</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0216" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/16</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/16 PCR m.scFv</td><td>152.6</td><td>1.86</td><td>2.10</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0216" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/16</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Matsumoto, Notsu, Wan</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/16 PCR m.scFv</td><td>4</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>11</td><td>30</td></tr>
 +
<tr><td>K592009</td><td>25</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>0</td><td>30</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0216" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/16</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td>5</td></tr>
 +
<tr><td>2</td><td>m.scFv 2/16 EPcut</td><td>30</td></tr>
 +
<tr><td>3</td><td>K592009 2/16EPcut</td><td>30</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/c/cc/T--Kyoto--2_16_No122_Gel_Extraction_Notsu_10_01_08_54_18.JPG">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0216" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/16</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>2/12 transformation J23108 colony3</td><td>LB(AMP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0217" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/17</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Uchino, Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Predenature</td><td>95℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>98℃</td><td>20sec</td><td>25</td></tr>
 +
<tr><td>Annealing</td><td>65℃</td><td>15sec</td><td>25</td></tr>
 +
<tr><td>Extension</td><td>72℃</td><td>30sec</td><td>25</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0217" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/17</h3></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
−
 
+
<span class="keln_researcher">Uchino, Kim</span>
−
<span class="keln_researcher">Sukegawa</span>
+
−
 
+
 
<table class="keln_table">
 
<table class="keln_table">
−
 
+
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
−
<tr><th>Templates/(μl)</th><th>Primer1/(μl)</th><th>Primer2/(μl)</th><th>Buffer/(μl)</th><th>MilliQ/(μl)</th><th>Any Other/(μl)</th><th>Total/(μl)</th></tr>
+
<tr><td>2/8 miniprep BAN-His14 /0.1</td><td>BAN-His-cex.Rv /0.75</td><td>VF /0.75</td><td>-</td><td>10.9</td><td>KAPA HiFi Ready Mix /12.5</td><td>25</td></tr>
−
 
+
<tr><td>2/8 miniprep BAN-His14 /0.1</td><td>BAN-His-clos.Rv /0.75</td><td>VF /0.75</td><td>-</td><td>10.9</td><td>KAPA HiFi Ready Mix /12.5</td><td>25</td></tr>
−
<tr><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
+
<tr><td>2/8 miniprep INP-His12 /0.1</td><td>INP-His-cex.Rv /0.75</td><td>VF /0.75</td><td>-</td><td>10.9</td><td>KAPA HiFi Ready Mix /12.5</td><td>25</td></tr>
−
 
+
<tr><td>2/8 miniprep INP-His12 /0.1</td><td>INP-His-clos.Rv /0.75</td><td>VF /0.75</td><td>-</td><td>10.9</td><td>KAPA HiFi Ready Mix /12.5</td><td>25</td></tr>
 
</table>
 
</table>
−
 
 
</div>
 
</div>
−
 
 
<!------------ Table END ------------>
 
<!------------ Table END ------------>
−
 
+
<!-- Table Generated by KELN Author: Notsu -->
−
<!-- Table Generated by KELN Author: Sukegawa -->
+
−
 
+
 
<div class="keln_container">
 
<div class="keln_container">
−
 
+
<a name="0217" class = "kyoto-jump"></a>
−
<a name="0101" class = "kyoto-jump"></a>
+
<span class="keln_date"><h3>2/17</h3></span>
−
 
+
<span class="keln_exp"><h4>Gel Extraction</h4></span>
−
<span class="keln_date"><h3>1/1</h3></span>
+
<span class="keln_researcher">Michimori, Wan, Yamamoto, Kim</span>
−
 
+
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>blank</td><td></td></tr>
 +
<tr><td>2</td><td>2/17PCR BAN-His-CBDcexRv+VF</td><td>25</td></tr>
 +
<tr><td>3</td><td>2/17PCR BAN-His-CBDclosRv+VF</td><td>25</td></tr>
 +
<tr><td>4</td><td>2/17PCR INP-His-CBDcexRv+VF</td><td>25</td></tr>
 +
<tr><td>5</td><td>2/17PCR INP-His-CBDclosRv+VF</td><td>25</td></tr>
 +
<tr><td>6</td><td>2/16PCR sf-GFP-CBDcex</td><td>50</td></tr>
 +
<tr><td>7</td><td>2/16PCR sf-GFP-CBDclos</td><td>50</td></tr>
 +
<tr><td>8</td><td>blank</td><td>-</td></tr>
 +
<tr><td>9</td><td>1kb ladder</td><td>-</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/2/2c/T--Kyoto--2_17_No8_Gel_Extraction_Notsu_10_02_07_09_31.2..jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0217" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/17</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Michimori, Wan, Yamamoto, Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/17PCR BAN-His-CBDcexRv+VF</td><td>10.0</td><td>1.86</td><td>0.43</td></tr>
 +
<tr><td>2/17PCR BAN-His-CBDclosRv+VF</td><td>34.6</td><td>1.91</td><td>1.81</td></tr>
 +
<tr><td>2/17PCR INP-His-CBDcexRv+VF</td><td>100.2</td><td>1.90</td><td>1.24</td></tr>
 +
<tr><td>2/17PCR INP-His-CBDclosRv+VF</td><td>116.5</td><td>1.90</td><td>2.08</td></tr>
 +
<tr><td>2/16PCR sf-GFP-CBDcex</td><td>244.0</td><td>1.91</td><td>2.08</td></tr>
 +
<tr><td>2/16PCR sf-GFP-CBDclos</td><td>208.9</td><td>1.90</td><td>2.01</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0217" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/17</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Predenature</td><td>95℃</td><td>3min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>98℃</td><td>20sec</td><td>35</td></tr>
 +
<tr><td>Annealing</td><td>65℃</td><td>15sec</td><td>35</td></tr>
 +
<tr><td>Extension</td><td>72℃</td><td>30sec</td><td>35</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0217" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/17</h3></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
−
 
+
<span class="keln_researcher">Yamamoto</span>
−
<span class="keln_researcher">Sukegawa</span>
+
−
 
+
 
<table class="keln_table">
 
<table class="keln_table">
−
 
+
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
−
<tr><th>Templates/(μl)</th><th>Primer1/(μl)</th><th>Primer2/(μl)</th><th>Buffer/(μl)</th><th>MilliQ/(μl)</th><th>Any Other/(μl)</th><th>Total/(μl)</th></tr>
+
<tr><td>2/17 Gel Extraction BAN-His-CBDcexRv+VF /8.5 + 3/17 Gel Extraction sf-GFP-CBDcex /0.5</td><td>VF /0.75</td><td>VR /0.75</td><td>-</td><td>2</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
−
 
+
<tr><td>2/17 Gel Extraction BAN-His-CBDclosRv+VF /7 + 3/17 Gel Extraction 2/16PCR sf-GFP-CBDclos /2</td><td>VF /0.75</td><td>VR /0.75</td><td>-</td><td>2</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
−
<tr><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
+
<tr><td>2/17 Gel Extraction INP-His-CBDcexRv+VF /6.5 + 3/17 Gel Extraction sf-GFP-CBDcex /1</td><td>VF /0.75</td><td>VR /0.75</td><td>-</td><td>3.5</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
−
 
+
<tr><td>2/17 Gel Extraction INP-His-CBDclosRv+VF /9 + 3/17 Gel Extraction sf-GFP-CBDclos /2</td><td>VF /0.75</td><td>VR /0.75</td><td>-</td><td>0</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
 
</table>
 
</table>
−
 
 
</div>
 
</div>
−
 
 
<!------------ Table END ------------>
 
<!------------ Table END ------------>
−
 
+
<!-- Table Generated by KELN Author: Notsu -->
−
<!-- Table Generated by KELN Author: Sukegawa -->
+
−
 
+
 
<div class="keln_container">
 
<div class="keln_container">
−
 
+
<a name="0217" class = "kyoto-jump"></a>
−
<a name="0101" class = "kyoto-jump"></a>
+
<span class="keln_date"><h3>2/17</h3></span>
−
 
+
<span class="keln_exp"><h4>Miniprep</h4></span>
−
<span class="keln_date"><h3>1/1</h3></span>
+
<span class="keln_researcher">Matsumoto, Kim</span>
−
 
+
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/16 liquid culture J23108 colony3</td><td>20.8</td><td>1.90</td><td>1.28</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0217" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/17</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Notsu, Wan</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/16 Gel Extraction m.scFv EPcut </td><td>14.3</td><td>1.87</td><td>1.09</td></tr>
 +
<tr><td>2/16 Gel Extraction K592009 EPcut</td><td>14.1</td><td>1.86</td><td>1.60</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0217" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/17</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Wan, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Vector/(&microl)</th><th>Insert/(&microl)</th><th>Ligation High/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/16 Gel Extraction K592009 EPcut /2.5</td><td>2/16 Gel Extraction m.scFv EPcut /5.0</td><td>7.5</td><td>15</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0217" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/17</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Wan, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>2/17 ligation m.scFv EPcut+K592009 EPcut</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0218" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/18</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/17 OE-PCR BAN-His-CBDcexRv+VF /8.5 + 3/17 Gel Extraction sf-GFP-CBDcex</td><td>72.7</td><td>1.87</td><td>1.93</td></tr>
 +
<tr><td>2/17 OE-PCR BAN-His-CBDclosRv+VF /7 + 3/17 Gel Extraction 2/16PCR sf-GFP-CBDclos</td><td>112.1</td><td>1.92</td><td>1.34</td></tr>
 +
<tr><td>2/17 OE-PCR INP-His-CBDcexRv+VF /6.5 + 3/17 Gel Extraction sf-GFP-CBDcex</td><td>102.4</td><td>1.89</td><td>1.96</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/a/ac/T--Kyoto--2_18_No11_Gel_Extraction%28Measurement%29_Notsu_10_02_07_28_40.1.jpg"><img src="https://static.igem.org/mediawiki/2016/7/7e/T--Kyoto--2_18_No11_Gel_Extraction%28Measurement%29_Notsu_10_02_07_28_40.2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0218" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/18</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/17 OE-PCR INP-His-CBDclosRv+VF /9 + 3/17 Gel Extraction sf-GFP-CBDclos</td><td>103.4</td><td>1.94</td><td>2.04</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/9/97/T--Kyoto--2_18_No11_Gel_Extraction%28Measurement%29_Notsu_10_02_07_32_29.1.jpg"><img src="https://static.igem.org/mediawiki/2016/3/30/T--Kyoto--2_18_No11_Gel_Extraction%28Measurement%29_Notsu_10_02_07_32_29.2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0218" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/18</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td>8</td></tr>
 +
<tr><td>2</td><td>2/17 OE-PCR BAN-His-CBDcexRv+VF /8.5 + 3/17 Gel Extraction sf-GFP-CBDcex</td><td>25</td></tr>
 +
<tr><td>3</td><td>2/17 OE-PCR BAN-His-CBDclosRv+VF /7 + 3/17 Gel Extraction 2/16PCR sf-GFP-CBDclos</td><td>25</td></tr>
 +
<tr><td>4</td><td>2/17 OE-PCR INP-His-CBDcexRv+VF /6.5 + 3/17 Gel Extraction sf-GFP-CBDcex</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0218" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/18</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td>8</td></tr>
 +
<tr><td>2</td><td>2/17 OE-PCR INP-His-CBDclosRv+VF /9 + 3/17 Gel Extraction sf-GFP-CBDclos</td><td>25</td></tr>
 +
<tr><td>3</td><td>blank</td><td>-</td></tr>
 +
<tr><td>4</td><td>blank</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0218" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/18</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher"></span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>BAN-CBDcex</td><td>72.3</td><td>1.85</td><td>1.87</td></tr>
 +
<tr><td>BAN-CBDclos</td><td>79.8</td><td>1.86</td><td>1.26</td></tr>
 +
<tr><td>INP-CBDcex</td><td>83.6</td><td>1.80</td><td>1.57</td></tr>
 +
<tr><td>INP-CBDclos</td><td>39.4</td><td>1.76</td><td>1.77</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0218" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/18</h3></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
−
 
+
<span class="keln_researcher">Kim</span>
−
<span class="keln_researcher">Sukegawa</span>
+
−
 
+
 
<table class="keln_table">
 
<table class="keln_table">
−
 
+
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
−
<tr><th>Templates/(μl)</th><th>Primer1/(μl)</th><th>Primer2/(μl)</th><th>Buffer/(μl)</th><th>MilliQ/(μl)</th><th>Any Other/(μl)</th><th>Total/(μl)</th></tr>
+
<tr><td>BAN-CBDcex /0.2</td><td>VF /1.25</td><td>VR /1.25</td><td>-</td><td>9.8</td><td>KAPA Pre Mix /12.5</td><td>25</td></tr>
−
 
+
<tr><td>BAN-CBDclos /0.2</td><td>VF /1.25</td><td>VR /1.25</td><td>-</td><td>9.8</td><td>KAPA Pre Mix /12.5</td><td>25</td></tr>
−
<tr><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
+
<tr><td>INP-CBDcex /0.2</td><td>VF /1.25</td><td>VR /1.25</td><td>-</td><td>9.8</td><td>KAPA Pre Mix /12.5</td><td>25</td></tr>
−
 
+
<tr><td>INP-CBDclos /0.2</td><td>VF /1.25</td><td>VR /1.25</td><td>-</td><td>9.8</td><td>KAPA Pre Mix /12.5</td><td>25</td></tr>
 
</table>
 
</table>
−
 
 
</div>
 
</div>
−
 
 
<!------------ Table END ------------>
 
<!------------ Table END ------------>
−
 
+
<!-- Table Generated by KELN Author: Notsu -->
−
<!-- Table Generated by KELN Author: Sukegawa -->
+
−
 
+
 
<div class="keln_container">
 
<div class="keln_container">
−
 
+
<a name="0218" class = "kyoto-jump"></a>
−
<a name="0101" class = "kyoto-jump"></a>
+
<span class="keln_date"><h3>2/18</h3></span>
−
 
+
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
−
<span class="keln_date"><h3>1/1</h3></span>
+
<span class="keln_researcher">Uchino</span>
−
 
+
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/17 transformation m.scFv EPcut+K592009 EPcut colony1</td><td>VF 1.25  VR 1.25</td><td>9.5</td><td>2×KAPA HiFi Fast Hotstart /12.5</td><td>25</td></tr>
 +
<tr><td>2/17 transformation m.scFv EPcut+K592009 EPcut colony2</td><td>VF 1.25  VR 1.25</td><td>9.5</td><td>2×KAPA HiFi Fast Hotstart /12.5</td><td>25</td></tr>
 +
<tr><td>2/17 transformation m.scFv EPcut+K592009 EPcut colony3</td><td>VF 1.25  VR 1.25</td><td>9.5</td><td>2×KAPA HiFi Fast Hotstart /12.5</td><td>25</td></tr>
 +
<tr><td>2/17 transformation m.scFv EPcut+K592009 EPcut colony4</td><td>VF 1.25  VR 1.25</td><td>9.5</td><td>2×KAPA HiFi Fast Hotstart /12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0218" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/18</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Predenature</td><td>95℃</td><td>2min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>95℃</td><td>10sec</td><td>30</td></tr>
 +
<tr><td>Annealing</td><td>55℃</td><td>15sec</td><td>30</td></tr>
 +
<tr><td>Extension</td><td>72℃</td><td>30sec</td><td>30</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0218" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/18</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Michimori, Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/18 PCR m.scFv EPcut+K592009 EPcut colony1</td><td>49.8</td><td>1.92</td><td>0.08</td></tr>
 +
<tr><td>2/18 PCR m.scFv EPcut+K592009 EPcut colony2</td><td>41.1</td><td>1.81</td><td>1.90</td></tr>
 +
<tr><td>2/18 PCR m.scFv EPcut+K592009 EPcut colony3</td><td>47.2</td><td>1.87</td><td>1.99</td></tr>
 +
<tr><td>2/18 PCR m.scFv EPcut+K592009 EPcut colony4</td><td>47.5</td><td>1.87</td><td>1.93</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0219" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/19</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>BAN-cex/(25)</td><td>161.1</td><td>1.88</td><td>2.13</td></tr>
 +
<tr><td>BAN-clos/(25)</td><td>217.9</td><td>1.86</td><td>2.10</td></tr>
 +
<tr><td>INP-cex/(25)</td><td>158.6</td><td>1.86</td><td>1.98</td></tr>
 +
<tr><td>INP-clos/(25)</td><td>163.6</td><td>1.85</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0219" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/19</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>BAN-cex</td><td>12.4</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H/(3)</td><td>0</td><td>12.6</td><td></td></tr>
 +
<tr><td>BAN-clos</td><td>9.2</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H/(3)</td><td>0</td><td>15.8</td><td></td></tr>
 +
<tr><td>INP-cex</td><td>12.6</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H/(3)</td><td>0</td><td>12.4</td><td></td></tr>
 +
<tr><td>INP-clos</td><td>12.2</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H/(3)</td><td>0</td><td>12.8</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yamamoto -->
 +
<div class="keln_container">
 +
<a name="0216” class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/16</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Kosaka</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>BAN-cex E, Pcut</td><td>30</td></tr>
 +
<tr><td>2</td><td>BAN-clos E, Pcut</td><td>30</td></tr>
 +
<tr><td>3</td><td>INP-cex E, Pcut</td><td>30</td></tr>
 +
<tr><td>0</td><td>1kb ladder</td><td>5</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/6/65/T--Kyoto--2_19_No18_Gel_Extraction_Kosaka_09_24_12_03_17.1.jpg"><img src="https://static.igem.org/mediawiki/2016/8/80/T--Kyoto--2_19_No18_Gel_Extraction_Kosaka_09_24_12_03_17.2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0219" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/19</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Egashira, Wan, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td>5</td></tr>
 +
<tr><td>2</td><td>2/19 Restriction Enzyme INP-clos EPcut</td><td>30</td></tr>
 +
<tr><td>3</td><td>2/19 Restriction Enzyme E1010 EPcut</td><td>30</td></tr>
 +
<tr><td>4</td><td>blank</td><td>-</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/6/6d/T--Kyoto--2_19_No18_Gel_Extraction_Notsu_10_15_08_27_59.1.jpg"><img src="https://static.igem.org/mediawiki/2016/b/b6/T--Kyoto--2_19_No18_Gel_Extraction_Notsu_10_15_08_27_59.2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: kosaka -->
 +
<div class="keln_container">
 +
<a name="0216" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/16</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Wan</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/19 Restriction Enzyme BAN-cex E, Pcut</td><td>9.6</td><td>1.85</td><td>0.84</td></tr>
 +
<tr><td>〃BAN-clos E, Pcut</td><td>18.9</td><td>1.83</td><td>0.71</td></tr>
 +
<tr><td>〃INP-cex E, Pcut</td><td>14.9</td><td>1.77</td><td>1.46</td></tr>
 +
<tr><td>〃INP-clos E, Pcut</td><td>13.4</td><td>1.73</td><td>1.67</td></tr>
 +
<tr><td>〃E1010 E, Pcut</td><td>19.6</td><td>1.86</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0219" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/19</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Wan, Egashira, Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>insertDNA/ul</td><td>VectoeDNA/ul</td><td>ligation high</td><td>total</td></tr>
 +
<tr><td>BAN-cex/4</td><td>E1010cpsb1c3/2</td><td>6</td><td>12</td></tr>
 +
<tr><td>BAN-clos/3</td><td>〃/3</td><td>6</td><td>12</td></tr>
 +
<tr><td>INP-cex/4.5</td><td>〃/1.5</td><td>6</td><td>12</td></tr>
 +
<tr><td>INP-clos/4.5</td><td>〃/1.5</td><td>6</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0219" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/19</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Wan</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>2/19 Ligation BAN-cex+E1010</td><td>10</td><td>DH5a/(20)</td><td>LB(CP)</td></tr>
 +
<tr><td>〃BAN-clos+E1010</td><td>10</td><td>DH5a/(20)</td><td>LB(CP)</td></tr>
 +
<tr><td>〃INP-cex+E1010</td><td>10</td><td>DH5a/(20)</td><td>LB(CP)</td></tr>
 +
<tr><td>〃INP-clos+E1010</td><td>10</td><td>DH5a/(20)</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0219" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/19</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Michimori, Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/18 purification m.scFv EPcut+K592009 EPcut colony1</td><td>12.5</td><td>0.5</td><td>0</td><td>0</td><td>0.5</td><td>10×H /1.5</td><td>0</td><td>0</td><td>15</td></tr>
 +
<tr><td>2/18 purification m.scFv EPcut+K592009 EPcut colony2</td><td>12.5</td><td>0.5</td><td>0</td><td>0</td><td>0.5</td><td>10×H /1.5</td><td>0</td><td>0</td><td>15</td></tr>
 +
<tr><td>2/18 purification m.scFv EPcut+K592009 EPcut colony3</td><td>12.5</td><td>0.5</td><td>0</td><td>0</td><td>0.5</td><td>10×H /1.5</td><td>0</td><td>0</td><td>15</td></tr>
 +
<tr><td>2/18 purification m.scFv EPcut+K592009 EPcut colony4</td><td>12.5</td><td>0.5</td><td>0</td><td>0</td><td>0.5</td><td>10×H /1.5</td><td>0</td><td>0</td><td>15</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0219" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/19</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1 </td><td>1kb ladder /5</td><td></td></tr>
 +
<tr><td>2</td><td>2/19 restricion enzyme digestion colony1 /10</td><td></td></tr>
 +
<tr><td>3</td><td>2/19 restricion enzyme digestion colony2 /10</td><td></td></tr>
 +
<tr><td>4</td><td>2/19 restricion enzyme digestion colony3 /10</td><td></td></tr>
 +
<tr><td>5</td><td>2/19 restricion enzyme digestion colony4 /10</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/2/24/T--Kyoto--2_19_No130_Electrophoresis_Notsu_10_01_09_25_34.JPG">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0219" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/19</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Egashira, Wan</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>2/17 transformaion m.scFv EPcut+K592009 EPcut colony2</td><td>LB(CP)</td></tr>
 +
<tr><td>2/17 transformaion m.scFv EPcut+K592009 EPcut colony3</td><td>LB(CP)</td></tr>
 +
<tr><td>2/17 transformaion m.scFv EPcut+K592009 EPcut colony4</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0219" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/19</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/19 transformation BAN-His-CBDcex colony1</td><td>vf 1.25 vr 1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>〃 colony2</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃 colony3</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>2/19 transformation BAN-His-CBDclos colony1</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃 colony2</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃 colony3</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>2/19 transformation INP-His-CBDcex colony1</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃 colony2</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃 colony3</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka ―>
 +
<div class="keln_container">
 +
<a name="0222" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/22</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>ladder 1kb/(5)</td><td>/</td></tr>
 +
<tr><td>2</td><td>2/22 PCR BAN-His-CBDcex colony1/(10)</td><td>515</td></tr>
 +
<tr><td>3</td><td>〃colony2/(10)</td><td>515</td></tr>
 +
<tr><td>4</td><td>〃colony3/(10)</td><td>515</td></tr>
 +
<tr><td>5</td><td>2/22 PCR BAN-His-CBDclos colony3/(10)</td><td>487</td></tr>
 +
<tr><td>6</td><td>2/22 PCR INP-His-CBDcex colony1/(10)</td><td>405</td></tr>
 +
<tr><td>7</td><td>〃colony2/(10)</td><td>405</td></tr>
 +
<tr><td>8</td><td>見えね</td><td></td></tr>
 +
<tr><td>9</td><td></td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/e/e5/T--Kyoto--2_22_No23_Electrophoresis_Kosaka_09_24_12_30_17.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0222" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/22</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>2/22 PCR BAN-His-cex colony1</td><td>CP/5ml</td></tr>
 +
<tr><td>〃colony2</td><td>〃</td></tr>
 +
<tr><td>〃colony3</td><td>〃</td></tr>
 +
<tr><td>2/22 PCR BAN-His-clos colony3</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0222" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/22</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Egashira, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/19 liquid culture m.scFv EPcut+K592009 EPcut colony2</td><td>137.4</td><td>1.89</td><td>2.12</td></tr>
 +
<tr><td>2/19 liquid culture m.scFv EPcut+K592009 EPcut colony3</td><td>164.0</td><td>1.89</td><td>2.09</td></tr>
 +
<tr><td>2/19 liquid culture m.scFv EPcut+K592009 EPcut colony4</td><td>138.1</td><td>1.86</td><td>2.16</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0223" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/19</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Wan, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>insertDNA/ul</td><td>VectoeDNA/ul</td><td>ligation high</td><td>total</td></tr>
 +
<tr><td>BAN-cex/4</td><td>E1010cpsb1c3/2</td><td>6</td><td>12</td></tr>
 +
<tr><td>BAN-clos/3</td><td>〃/3</td><td>6</td><td>12</td></tr>
 +
<tr><td>INP-cex/4.5</td><td>〃/1.5</td><td>6</td><td>12</td></tr>
 +
<tr><td>INP-clos/4.5</td><td>〃/1.5</td><td>6</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0223" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/23</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Wan</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>2/23 Ligation BAN-cex+E1010</td><td>2</td><td>DH5a/(20)</td><td>LB(CP)</td></tr>
 +
<tr><td>〃clos〃</td><td>2</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃INP-cex〃</td><td>2</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃INP-clos〃</td><td>2</td><td>〃</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0223" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/23</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/23 transformation BAN-His-CBDclos colony1</td><td>vf1.25 vr1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>〃colony2</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony3</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>2/23 transformation INP-His-CBDcex colony1</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony2</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony3</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>2/23 transformation INP-His-CBDclos colony1</td><td>〃</td><td>〃</td><td>〃</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0224" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/24</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/22 liqid culture BAN-Hi-cex colony1</td><td>45.3</td><td>1.88</td><td>2.08</td></tr>
 +
<tr><td>〃colony2</td><td>47.1</td><td>1.89</td><td>1.98</td></tr>
 +
<tr><td>〃colony3</td><td>43.8</td><td>1.96</td><td>2.08</td></tr>
 +
<tr><td>2/22 BAN-His-clos colony3</td><td>53.7</td><td>1.82</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0224" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/24</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>2/23 transformation BAN-clos colony1</td><td>LB(CP/5ml)</td></tr>
 +
<tr><td>〃colony2</td><td>〃</td></tr>
 +
<tr><td>〃colony3</td><td>〃</td></tr>
 +
<tr><td>2/23 transformation INP-cex colony2</td><td>〃</td></tr>
 +
<tr><td>〃colony3</td><td>〃</td></tr>
 +
<tr><td>2/23 transformation INP-clos colony1</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0225" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/25</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Egashira, Kim</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/23 transformation BAN-His-CBDclos colony1</td><td>79.9</td><td>1.89</td><td>2.15</td></tr>
 +
<tr><td>2/23 transformation BAN-His-CBDclos colony2</td><td>68.0</td><td>1.88</td><td>2.02</td></tr>
 +
<tr><td>2/23 transformation BAN-His-CBDclos colony3</td><td>80.7</td><td>1.94</td><td>2.18</td></tr>
 +
<tr><td>2/23 transformation INP-His-CBDcex colony2</td><td>106.0</td><td>1.91</td><td>2.17</td></tr>
 +
<tr><td>2/23 transformation INP-His-CBDcex colony3</td><td>136.7</td><td>1.93</td><td>2.23</td></tr>
 +
<tr><td>2/23 transformation INP-His-CBDclos colony1</td><td>183.7</td><td>1.91</td><td>2.21</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0225" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/25</h3></span>
 +
<span class="keln_exp"><h4>Sequence</h4></span>
 +
<span class="keln_researcher">Michimori</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA template/(&microl)</th><th>Primer/(&microl)</th><th>MilliQ(&microl)</th><th>Total(&microl)</th></tr>
 +
<tr><td>Bcx 1-F</td><td>2/24 miniprep BAN-His-CBDcex colony1 /6.6</td><td>VF /0.2</td><td>3.2</td><td>10</td></tr>
 +
<tr><td>Bcx 1-R</td><td>2/24 miniprep BAN-His-CBDcex colony1 /6.6</td><td>VR /0.2</td><td>3.2</td><td>10</td></tr>
 +
<tr><td>Bcx 2-F</td><td>2/24 miniprep BAN-His-CBDcex colony2 /6.4</td><td>VF /0.2</td><td>3.4</td><td>10</td></tr>
 +
<tr><td>Bcx 2-R</td><td>2/24 miniprep BAN-His-CBDcex colony2 /6.4</td><td>VR /0.2</td><td>3.4</td><td>10</td></tr>
 +
<tr><td>Bcl1-F</td><td>2/25 miniprep BAN-His-CBDclos colony1 /3.8</td><td>VF /0.2</td><td>6.0</td><td>10</td></tr>
 +
<tr><td>Bcl1-R</td><td>2/25 miniprep BAN-His-CBDclos colony1 /3.8</td><td>VR /0.2</td><td>6.0</td><td>10</td></tr>
 +
<tr><td>Bcl2-F</td><td>2/25 miniprep BAN-His-CBDclos colony3 /3.7</td><td>VF /0.2</td><td>6.1</td><td>10</td></tr>
 +
<tr><td>Bcl2-R</td><td>2/25 miniprep BAN-His-CBDclos colony3 /3.7</td><td>VR /0.2</td><td>6.1</td><td>10</td></tr>
 +
<tr><td>Icx1-F</td><td>2/25 miniprep INP-His-CBDcex colony2 /2.8</td><td>VF /0.2</td><td>7.0</td><td>10</td></tr>
 +
<tr><td>Icx1-R</td><td>2/25 miniprep INP-His-CBDcex colony2 /2.8</td><td>VR /0.2</td><td>7.0</td><td>10</td></tr>
 +
<tr><td>Icx2-F</td><td>2/25 miniprep INP-His-CBDcex colony3 /2.2</td><td>VF /0.2</td><td>7.6</td><td>10</td></tr>
 +
<tr><td>Icx2-R</td><td>2/25 miniprep INP-His-CBDcex colony3 /2.2</td><td>VR /0.2</td><td>7.6</td><td>10</td></tr>
 +
<tr><td>Icl1-F</td><td>2/25 miniprep INP-His-CBDclos colony1 /1.6</td><td>VF /0.2</td><td>8.2</td><td>10</td></tr>
 +
<tr><td>Icl1-R</td><td>2/25 miniprep INP-His-CBDclos colony1 /1.6</td><td>VR /0.2</td><td>8.2</td><td>10</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Koide -->
 +
<div class="keln_container">
 +
<a name="0225" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/25</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>2/19 transformation BAN-His-cex colony1</td><td>LB(CP/5ml)</td></tr>
 +
<tr><td>2/19 transformation BAN-His-cex colony2</td><td>LB(CP/5ml)</td></tr>
 +
<tr><td>2/23 transformation BAN-His-clos colony1</td><td>LB(CP/5ml)</td></tr>
 +
<tr><td>2/23 transformation BAN-His-clos colony3</td><td>LB(CP/5ml)</td></tr>
 +
<tr><td>2/23 transformaation INP-His-cex colony2</td><td>LB(CP/5ml)</td></tr>
 +
<tr><td>2/23 transformation INP-His-cex colony3</td><td>LB(CP/5ml)</td></tr>
 +
<tr><td>INP-His-clos colony1</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0225" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/25</h3></span>
 +
<span class="keln_exp"><h4>Sequence</h4></span>
 +
<span class="keln_researcher">Michimori</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA template/(&microl)</th><th>Primer/(&microl)</th><th>MilliQ(&microl)</th><th>Total(&microl)</th></tr>
 +
<tr><td>S2-F</td><td>2/22 miniprep scFv colony2 /2.2</td><td>VF /0.2</td><td>7.6</td><td>10</td></tr>
 +
<tr><td>S2-R</td><td>2/22 miniprep scFv colony2 /2.2</td><td>VR /0.2</td><td>7.6</td><td>10</td></tr>
 +
<tr><td>S3-F</td><td>2/22 miniprep scFv colony3 /1.8</td><td>VF /0.2</td><td>8.0</td><td>10</td></tr>
 +
<tr><td>S3-R</td><td>2/22 miniprep scFv colony3 /1.8</td><td>VR /0.2</td><td>8.0</td><td>10</td></tr>
 +
<tr><td>S4-F</td><td>2/22 miniprep scFv colony4 /2.2</td><td>VF /0.2</td><td>7.6</td><td>10</td></tr>
 +
<tr><td>S4-R</td><td>2/22 miniprep scFv colony4 /2.2</td><td>VR /0.2</td><td>7.6</td><td>10</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note"> 114C-R</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0225" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/25</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Egashira, Kim, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>2/17 transformaion m.scFv EPcut+K592009 EPcut colony2</td><td>LB(CP)</td></tr>
 +
<tr><td>2/17 transformaion m.scFv EPcut+K592009 EPcut colony3</td><td>LB(CP)</td></tr>
 +
<tr><td>2/17 transformaion m.scFv EPcut+K592009 EPcut colony4</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Iguchi -->
 +
<div class="keln_container">
 +
<a name="0226" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/26</h3></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
−
 
+
<span class="keln_researcher">Li</span>
−
<span class="keln_researcher">Sukegawa</span>
+
−
 
+
 
<table class="keln_table">
 
<table class="keln_table">
−
 
+
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
−
<tr><th>Templates/(μl)</th><th>Primer1/(μl)</th><th>Primer2/(μl)</th><th>Buffer/(μl)</th><th>MilliQ/(μl)</th><th>Any Other/(μl)</th><th>Total/(μl)</th></tr>
+
<tr><td>J23100</td><td>VF/ 0.75</td><td>J23100+RBSRv</td><td></td><td>11</td><td>PrimeSTARMax Premix/ 12.5</td><td>25</td></tr>
−
 
+
<tr><td>J23108</td><td>VF/ 0.75</td><td>J23108+RBSRv</td><td></td><td>11</td><td>PrimeSTARMax Premix/ 12.5</td><td>25</td></tr>
−
<tr><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
+
<tr><td>J23114</td><td>VF/ 0.75</td><td>J23114+RBSRv</td><td></td><td>11</td><td></td><td>25</td></tr>
−
 
+
 
</table>
 
</table>
−
 
 
</div>
 
</div>
−
 
 
<!------------ Table END ------------>
 
<!------------ Table END ------------>
−
 
+
<!-- Table Generated by KELN Author: Notsu -->
−
<!-- Table Generated by KELN Author: Sukegawa -->
+
−
 
+
 
<div class="keln_container">
 
<div class="keln_container">
−
 
+
<a name="0226" class = "kyoto-jump"></a>
−
<a name="0101" class = "kyoto-jump"></a>
+
<span class="keln_date"><h3>2/26</h3></span>
−
 
+
−
<span class="keln_date"><h3>1/1</h3></span>
+
−
 
+
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
−
 
+
<span class="keln_researcher">Li</span>
−
<span class="keln_researcher">Sukegawa</span>
+
−
 
+
 
<table class="keln_table">
 
<table class="keln_table">
−
 
+
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
−
<tr><th>Templates/(μl)</th><th>Primer1/(μl)</th><th>Primer2/(μl)</th><th>Buffer/(μl)</th><th>MilliQ/(μl)</th><th>Any Other/(μl)</th><th>Total/(μl)</th></tr>
+
<tr><td>J23100 iGEM Part</td><td>VF /0.75</td><td>J23100+RBS Rv /0.75</td><td>-</td><td>11</td><td>PrimeSTAR Max Premix /12.5</td><td>25</td></tr>
−
 
+
<tr><td>J23108 iGEM Part</td><td>VF /0.75</td><td>J23108+RBS Rv /0.75</td><td>-</td><td>11</td><td>PrimeSTAR Max Premix /12.5</td><td>25</td></tr>
−
<tr><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
+
<tr><td>J23114 iGEM Part</td><td>VF /0.75</td><td>J23114+RBS Rv /0.75</td><td>-</td><td>11</td><td>PrimeSTAR Max Premix /12.5</td><td>25</td></tr>
−
 
+
 
</table>
 
</table>
−
 
 
</div>
 
</div>
−
 
 
<!------------ Table END ------------>
 
<!------------ Table END ------------>
−
 
+
<!-- Table Generated by KELN Author: Notsu -->
−
<!-- Table Generated by KELN Author: Sukegawa -->
+
−
 
+
 
<div class="keln_container">
 
<div class="keln_container">
−
 
+
<a name="0226" class = "kyoto-jump"></a>
−
<a name="0101" class = "kyoto-jump"></a>
+
<span class="keln_date"><h3>2/26</h3></span>
−
 
+
<span class="keln_exp"><h4>Electrophoresis</h4></span>
−
<span class="keln_date"><h3>1/1</h3></span>
+
<span class="keln_researcher">Egashira</span>
−
 
+
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /5</td><td>-</td></tr>
 +
<tr><td>2</td><td>2/26 PCR J23100 /5</td><td>193+α</td></tr>
 +
<tr><td>3</td><td>2/26 PCR J23108 /5</td><td>193+α</td></tr>
 +
<tr><td>4</td><td>2/26 PCR J23114 /5</td><td>193+α</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0226" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/26</h3></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
−
 
+
<span class="keln_researcher">Egashira</span>
−
<span class="keln_researcher">Sukegawa</span>
+
−
 
+
 
<table class="keln_table">
 
<table class="keln_table">
−
 
+
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
−
<tr><th>Templates/(μl)</th><th>Primer1/(μl)</th><th>Primer2/(μl)</th><th>Buffer/(μl)</th><th>MilliQ/(μl)</th><th>Any Other/(μl)</th><th>Total/(μl)</th></tr>
+
<tr><td>J23100 iGEM Part /0.1</td><td>VF 0.75</td><td>J23100+RBS Rv /0.75</td><td>-</td><td>10.9</td><td>KAPA HiFi /12.5</td><td>25</td></tr>
−
 
+
<tr><td>J23108 iGEM Part /0.1</td><td>VF 0.75</td><td>J23108+RBS Rv /0.75</td><td>-</td><td>10.9</td><td>KAPA HiFi /12.5</td><td>25</td></tr>
−
<tr><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
+
<tr><td>J23114 iGEM Part /0.1</td><td>VF 0.75</td><td>J23114+RBS Rv /0.75</td><td>-</td><td>10.9</td><td>KAPA HiFi /12.5</td><td>25</td></tr>
−
 
+
 
</table>
 
</table>
−
 
+
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0226" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/26</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder </td><td>5</td></tr>
 +
<tr><td>2</td><td>2/26 PCR J23100</td><td>25</td></tr>
 +
<tr><td>3</td><td>2/26 PCR J23108</td><td>25</td></tr>
 +
<tr><td>4</td><td>2/26 PCR J23114</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0229" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/29</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Egashira, Wan</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/26 PCR J23100</td><td>1.5</td><td>2.71</td><td>0.09</td></tr>
 +
<tr><td>2/26 PCR J23108</td><td>3.6</td><td>18.64</td><td>0.01</td></tr>
 +
<tr><td>2/26 PCR J23114</td><td>1.7</td><td>3.05</td><td>0.13</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0229" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>2/29</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Wan, Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>J23100 iGEM Part /0.2</td><td>VF /1.5</td><td>J23100+RBS Rv /1.5</td><td>-</td><td>21.8</td><td>KAPA HiFi /25</td><td>50</td></tr>
 +
<tr><td>J23108 iGEM Part /0.2</td><td>VF /1.5</td><td>J23108+RBS Rv /1.5</td><td>-</td><td>21.8</td><td>KAPA HiFi /25</td><td>50</td></tr>
 +
<tr><td>J23114 iGEM Part /0.2</td><td>VF /1.5</td><td>J23114+RBS Rv /1.5</td><td>-</td><td>21.8</td><td>KAPA HiFi /25</td><td>50</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0301" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/1</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /5</td><td>-</td></tr>
 +
<tr><td>2</td><td>3/1 PCR J23100 /5</td><td>about 200</td></tr>
 +
<tr><td>3</td><td>3/1 PCR J23108 /5</td><td>about 200</td></tr>
 +
<tr><td>4</td><td>3/1 PCR J23114 /5</td><td>about 200</td></tr>
 +
<tr><td>5</td><td>3/1 PCR m.scFv colony3 /5</td><td>949</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0301" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/1</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Kim, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>100bp ladder /5</td><td>-</td></tr>
 +
<tr><td>2</td><td>3/1 OE-PCR J23100 + m.scFv colony3 /2</td><td>about 1200</td></tr>
 +
<tr><td>3</td><td>3/1 OE-PCR J23108 + m.scFv colony3 /2</td><td>about 1200</td></tr>
 +
<tr><td>4</td><td>3/1 OE-PCR J23114 + m.scFv colony3 /2</td><td>about 1200</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0301" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/1</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Kim, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>3/1 OE-PCR J23100 + m.scFv colony3 </td><td>180.5</td><td>1.89</td><td>1.83</td></tr>
 +
<tr><td>3/1 OE-PCR J23108 + m.scFv colony3</td><td>183.3</td><td>1.87</td><td>1.96</td></tr>
 +
<tr><td>3/1 OE-PCR J23114 + m.scFv colony3</td><td>178.8</td><td>1.87</td><td>2.08</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0301" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/1</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Kim, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/1 PCR J23100 /1 + 3/1 PCR m.scFv colony3 /0.5</td><td>VF /0.5</td><td>VR /0.75</td><td>-</td><td>9.5</td><td>Prime Star Premix /12.5</td><td>25</td></tr>
 +
<tr><td>3/1 PCR J23108 /1 + 3/1 PCR m.scFv colony3 /0.5</td><td>VF /0.5</td><td>VR /0.75</td><td>-</td><td>9.5</td><td>Prime Star Premix /12.5</td><td>25</td></tr>
 +
<tr><td>3/1 PCR J23114 /2 + 3/1 PCR J23114 /0.5</td><td>VF /0.5</td><td>VR /0.75</td><td>-</td><td>8.5</td><td>Prime Star Premix /12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0301" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/1</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Kim, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td></td></tr>
 +
<tr><td>2</td><td>3/1 EPcut J23114 + m.scFv</td><td>20</td></tr>
 +
<tr><td>3</td><td>3/1 EPcut J23108 + m.scFv</td><td>20</td></tr>
 +
<tr><td>4</td><td>3/1 EPcut J23100 + m.scFv</td><td>20</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0301" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/1</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Li, Kim, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/1 OE-PCR J23100 + m.scFv colony3</td><td>11</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>-</td><td>14</td><td>30</td></tr>
 +
<tr><td>3/1 OE-PCR J23108 + m.scFv colony3</td><td>11</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>-</td><td>14</td><td>30</td></tr>
 +
<tr><td>3/1 OE-PCR J23114 + m.scFv colony3</td><td>11</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>-</td><td>14</td><td>30</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0302" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/2</h3></span>
 +
<span class="keln_exp"><h4>PartsAwaking</h4></span>
 +
<span class="keln_researcher">Michimori, Kim, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Description</th><th>PartName</th><th>Backbone</th><th>Well</th><th>PlateYear</th><th>PlateNumber</th><th>MilliQ(&microl)</th></tr>
 +
<tr><td>f1 origin</td><td>K314110</td><td>pSB1C3</td><td>1A</td><td>2016</td><td>1</td><td>10</td></tr>
 +
<tr><td>T1 terminator from E. coli rrnB</td><td>K731722</td><td>pSB1C3</td><td>1B</td><td>2016</td><td>1</td><td>10</td></tr>
 +
<tr><td>Promoter of the CaiF protein</td><td>K398326</td><td>pSB1C3</td><td>1C</td><td>2016</td><td>1</td><td>10</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0302" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/2</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Michimori</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>3/2 Parts Awakening K314110</td><td>10</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>3/2 Parts Awakening K731722</td><td>10</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>3/2 Parts Awakening K398326</td><td>10</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0302" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/2</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>J23100+m.scFv 3/1 EPcut </td><td>17.7</td><td>1.90</td><td>1.68</td></tr>
 +
<tr><td>J23108+m.scFv 3/1 EPcut </td><td>11.9</td><td>2.04</td><td>0.26</td></tr>
 +
<tr><td>J23114+m.scFv 3/1 EPcut </td><td>16.8</td><td>1.97</td><td>1.79</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0302" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/2</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Wan</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>E0020</td><td>40</td><td>2</td><td>0</td><td>0</td><td>2</td><td>10×H /6</td><td>-</td><td>10</td><td>60</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0302" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/2</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>J23100+pSB1C3</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>J23108+pSB1C3</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>J23114+pSB1C3</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0303" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/3</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/9 miniprep E1010 /1</td><td>VF 0.75</td><td>VR 0.75</td><td>-</td><td>10</td><td>Prime Star premix 12.5</td><td>25</td></tr>
 +
<tr><td>2/24 miniprep BAN-His-CBDcex colony2 /1</td><td>VF 0.75</td><td>VR 0.75</td><td>-</td><td>10</td><td>Prime Star premix 12.5</td><td>25</td></tr>
 +
<tr><td>2/25 miniprep BAN-His-CBDclos /1</td><td>VF 0.75</td><td>VR 0.75</td><td>-</td><td>10</td><td>Prime Star premix 12.5</td><td>25</td></tr>
 +
<tr><td>2/25 miniprep INP-His-CBDcex /1</td><td>VF 0.75</td><td>VR 0.75</td><td>-</td><td>10</td><td>Prime Star premix 12.5</td><td>25</td></tr>
 +
<tr><td>2/25 miniprep INP-His-CBDclos /1</td><td>VF 0.75</td><td>VR 0.75</td><td>-</td><td>10</td><td>Prime Star premix 12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0303" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/3</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>3/2 EPcut E0020</td><td>23.1</td><td>1.85</td><td>1.69</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0303" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/3</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td></td></tr>
 +
<tr><td>2</td><td>3/2 EPcut E0020 (pSB1C3)</td><td>20</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0303" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/3</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Wan</span>
 +
<table class="keln_table">
 +
<tr><th>Vector/(&microl)</th><th>Insert/(&microl)</th><th>Ligation High/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/3 Gel Extraction E0020 EPcut /4</td><td>3/2 Gel Extraction J23100+m.scFv EPcut /12</td><td>16</td><td>32</td></tr>
 +
<tr><td>3/3 Gel Extraction E0020 EPcut /4</td><td>3/2 Gel Extraction J23108+m.scFv EPcut /12</td><td>20</td><td>40</td></tr>
 +
<tr><td>3/3 Gel Extraction E0020 EPcut /4</td><td>3/2 Gel Extraction J23114+m.scFv EPcut /12</td><td>16</td><td>32</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Iguchi -->
 +
<div class="keln_container">
 +
<a name="0303" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/3</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Wan, Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>J23100+PSB1C3</td><td>2</td><td>DH5alpha/ 20</td><td>LB(CP)</td></tr>
 +
<tr><td>J23108+PSB1C3</td><td>2</td><td>DH5alpha/ 20</td><td>LB(CP)</td></tr>
 +
<tr><td>J23114+PSB1C3</td><td>2</td><td>DH5alpha</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0303" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/3</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Wan, Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>K314110 colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>K731722 colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>K398326 colony1</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0304" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/4</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Li, Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>J23100+pSB1C3</td><td>10</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>J23108+pSB1C3</td><td>10</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>J23114+pSB1C3</td><td>10</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0307" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/7</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/25 liquid culture BAN-His-CBDcex colony2</td><td>71.4</td><td>1.83</td><td>1.43</td></tr>
 +
<tr><td>2/25 liquid culture BAN-His-CBDclos colony1</td><td>61.6</td><td>1.83</td><td>1.90</td></tr>
 +
<tr><td>2/25 liquid culture INP-His-CBDcex colony3</td><td>83.4</td><td>1.94</td><td>1.92</td></tr>
 +
<tr><td>2/25 liquid culture INP-His-CBDclos colony1</td><td>100.7</td><td>1.87</td><td>1.44</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0307" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/7</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Wan</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>3/7 miniprep BAN-His-CBDcex</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>3/7 miniprep BAN-His-CBDclos</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>3/7 miniprep INP-His-CBDcex</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>3/7 miniprep INP-His-CBDclos</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td> </td><td></td><td></td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0307" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/7</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>3/3 ligation J23100+pSB1C3</td><td></td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>3/3 ligation J23108+pSB1C3</td><td></td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>3/3 ligation J23114+pSB1C3</td><td></td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0307" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/7</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/4 transformtion J23100 colony1</td><td>VF 1.25    VR 1.25</td><td>10</td><td>KAPA2G /12.5</td><td>25</td></tr>
 +
<tr><td>3/4 transformtion J23100 colony2</td><td>VF 1.25    VR 1.25</td><td>10</td><td>KAPA2G /12.5</td><td>25</td></tr>
 +
<tr><td>3/4 transformtion J23108 colony1</td><td>VF 1.25    VR 1.25</td><td>10</td><td>KAPA2G /12.5</td><td>25</td></tr>
 +
<tr><td>3/4 transformtion J23108 colony2</td><td>VF 1.25    VR 1.25</td><td>10</td><td>KAPA2G /12.5</td><td>25</td></tr>
 +
<tr><td>3/4 transformtion J23114 colony1</td><td>VF 1.25    VR 1.25</td><td>10</td><td>KAPA2G /12.5</td><td>25</td></tr>
 +
<tr><td>3/4 transformtion J23114 colony2</td><td>VF 1.25    VR 1.25</td><td>10</td><td>KAPA2G /12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0307" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/7</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Predenature</td><td>95℃</td><td>2min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>95℃</td><td>15sec</td><td>35</td></tr>
 +
<tr><td>Annealing</td><td>55℃</td><td>15sec</td><td>35</td></tr>
 +
<tr><td>Extension</td><td>72℃</td><td>5sec</td><td>35</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>10min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0307" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/7</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Wan, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /5</td><td></td></tr>
 +
<tr><td>2</td><td>3/7 colony PCR J23100 colony1 /10</td><td>1000+α</td></tr>
 +
<tr><td>3</td><td>3/7 colony PCR J23100 colony2 /10</td><td>1000+α</td></tr>
 +
<tr><td>4</td><td>3/7 colony PCR J23108 colony1 /10</td><td>1000+α</td></tr>
 +
<tr><td>5</td><td>3/7 colony PCR J23108 colony2 /10</td><td>1000+α</td></tr>
 +
<tr><td>6</td><td>3/7 colony PCR J23114 colony1 /10</td><td>1000+α</td></tr>
 +
<tr><td>7</td><td>3/7 colony PCR J23114 colony2 /10</td><td>1000+α</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/c/c9/T--Kyoto--3_7_No154_Electrophoresis_Notsu_10_02_10_08_42.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0307" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/7</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Predenature</td><td>95℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>98℃</td><td>20sec</td><td>35</td></tr>
 +
<tr><td>Annealing</td><td>60℃</td><td>15sec</td><td>35</td></tr>
 +
<tr><td>Extension</td><td>72℃</td><td>5min</td><td>35</td></tr>
 +
<tr><td>Final Elongation</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
<tr><td>Final Holding</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0307" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/7</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/1 purification J23100</td><td>VF /3</td><td>VR /3</td><td>-</td><td>44</td><td>KAPA HiFi /50</td><td>100</td></tr>
 +
<tr><td>3/1 purification J23100</td><td>VF /3</td><td>VR /3</td><td>-</td><td>44</td><td>KAPA HiFi /50</td><td>100</td></tr>
 +
<tr><td>3/1 purification J23108</td><td>VF /3</td><td>VR /3</td><td>-</td><td>44</td><td>KAPA HiFi /50</td><td>100</td></tr>
 +
<tr><td>3/1 purification J23108</td><td>VF /3</td><td>VR /3</td><td>-</td><td>44</td><td>KAPA HiFi /50</td><td>100</td></tr>
 +
<tr><td>3/1 purification J23114</td><td>VF /3</td><td>VR /3</td><td>-</td><td>44</td><td>KAPA HiFi /50</td><td>100</td></tr>
 +
<tr><td>3/1 purification J23114</td><td>VF /3</td><td>VR /3</td><td>-</td><td>44</td><td>KAPA HiFi /50</td><td>100</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0307" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/7</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/7 transformation BAN-CBDcex colony1</td><td>VF  1.5  VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>3/7 transformation BAN-cex colony2</td><td>VF  1.5  VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>3/7 transformation BAN-clos colony1</td><td>VF  1.5  VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>3/7 transformation BAN-closcolony1</td><td>VF  1.5  VR1.25</td><td>10</td><td>12.5</td><td></td></tr>
 +
<tr><td>3/7 transformation INP-cex colony2</td><td>VF  1.5  VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>3/7 transformation  INP-cex colony2</td><td>VF  1.5  VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>3/7 transformation INP-clos colony1</td><td>VF  1.5  VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>3/7 transformation INP-clos colony 2</td><td>VF  1.5  VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0308" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/8</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>3/7 PCR J23100</td><td>423.3</td><td>1.84</td><td>2.20</td></tr>
 +
<tr><td>3/7 PCR J23108</td><td>439.1</td><td>1.85</td><td>2.11</td></tr>
 +
<tr><td>3/7 PCR J23114</td><td>451.2</td><td>1.81</td><td>1.91</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0308" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/8</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/8 purification J23100</td><td>5</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>20</td><td>30</td></tr>
 +
<tr><td>3/8 purification J23108</td><td>5</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>20</td><td>30</td></tr>
 +
<tr><td>3/8 purification J23114</td><td>5</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>20</td><td>30</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0308" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/8</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Li, Kim, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>J23114 3/8 EPcut</td><td>13.7</td><td>3.62</td><td>0.03</td></tr>
 +
<tr><td>J23108 3/8 EPcut</td><td>15.4</td><td>1.83</td><td>1.14</td></tr>
 +
<tr><td>J23100 3/8 EPcut</td><td>14.5</td><td>1.79</td><td>1.03</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0309" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/9</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>K731722</td><td>67.2</td><td>1.70</td><td>1.02</td></tr>
 +
<tr><td>K314110</td><td>28.9</td><td>1.86</td><td>1.14</td></tr>
 +
<tr><td>K398326</td><td>95.6</td><td>1.64</td><td>0.94</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0309" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/9</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/1 miniprep K314110</td><td>25</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>0</td><td>30</td></tr>
 +
<tr><td>3/1 miniprep K731722</td><td>25</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>0</td><td>30</td></tr>
 +
<tr><td>3/1 miniprep K398326</td><td>25</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>0</td><td>30</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0309" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/9</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Li</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/8 purification J23100</td><td>5</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>20</td><td>30</td></tr>
 +
<tr><td>3/8 purification J23108</td><td>5</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>20</td><td>30</td></tr>
 +
<tr><td>3/8 purification J23114</td><td>5</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>20</td><td>30</td></tr>
 +
<tr><td>3/8 purification J23100</td><td>20</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>5</td><td>30</td></tr>
 +
<tr><td>3/8 purification J23108</td><td>20</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>5</td><td>30</td></tr>
 +
<tr><td>3/8 purification J23114</td><td>20</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>5</td><td>30</td></tr>
 +
<tr><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td>30</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0311" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/11</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td>5</td></tr>
 +
<tr><td>2</td><td>3/10 E.P. cut J23100+scFv</td><td>15</td></tr>
 +
<tr><td>3</td><td>3/10 E.P. cut J23100+scFv</td><td>15</td></tr>
 +
<tr><td>4</td><td>3/10 E.P.cut J23108+scFv</td><td>15</td></tr>
 +
<tr><td>5</td><td>3/10 E.P.cut J23108+scFv</td><td>15</td></tr>
 +
<tr><td>6</td><td>3/10 E.P.cut J23114+scFv</td><td>15</td></tr>
 +
<tr><td>7</td><td>3/10 E.P.cut J23114+scFv</td><td>15</td></tr>
 +
<tr><td>8</td><td>3/9 E.P. cut J23100+scFv</td><td>15</td></tr>
 +
<tr><td>9</td><td>3/9 E.P. cut J23100+scFv</td><td>15</td></tr>
 +
<tr><td>10</td><td>3/9 Ｅ.P. cut J23119 scFv</td><td>15</td></tr>
 +
<tr><td>11</td><td>3/9 Ｅ.P. cut J23119 scFv</td><td>15</td></tr>
 +
<tr><td>12</td><td>3/9 E.P.cut J23114+scFv</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0311" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/11</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kbp ladder</td><td>15</td></tr>
 +
<tr><td>2</td><td>3/9 E.P. cut K314110</td><td>15</td></tr>
 +
<tr><td>3</td><td>3/9 E.P. cut K314110</td><td>15</td></tr>
 +
<tr><td>4</td><td>3/9 E.P cut  K731722</td><td>15</td></tr>
 +
<tr><td>5</td><td>3/9 E.P cut  K731722</td><td>15</td></tr>
 +
<tr><td>6</td><td>3/9 E.P cut K7398326</td><td>15</td></tr>
 +
<tr><td>7</td><td>3/9 E.P cut K7398326</td><td>15</td></tr>
 +
<tr><td>8</td><td>3/9 E.P> cut J23114+scFv</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/9/9a/T--Kyoto--3_11_No164_Gel_Extraction_Yoshimoto_09_16_09_36_49%2C1.jpg"> <img src="https://static.igem.org/mediawiki/2016/8/85/T--Kyoto--3_11_No164_Gel_Extraction_Yoshimoto_09_16_09_36_49%2C2.jpg"> <img src="https://static.igem.org/mediawiki/2016/8/8c/T--Kyoto--3_11_No164_Gel_Extraction_Yoshimoto_09_16_09_36_49%2C3.jpg"> <img src="https://static.igem.org/mediawiki/2016/c/ce/T--Kyoto--3_11_No164_Gel_Extraction_Yoshimoto_09_16_09_36_49%2C4.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0311" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/11</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Yoshimoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>3/11 Gel Ex J23108+scFv</td><td>18.9</td><td>1.90</td><td>0.19</td></tr>
 +
<tr><td>3/11 Gel Ex J23108+scFv</td><td>7.0</td><td>1.92</td><td>0.57</td></tr>
 +
<tr><td>3/11 Gel Ex J23114+scFv</td><td>5.3</td><td>2.37</td><td>0.25</td></tr>
 +
<tr><td>3/11 Gel Ex J23100+scFv</td><td>39.2</td><td>1.93</td><td>1.29</td></tr>
 +
<tr><td>3/11 Gel Ex J23108+scFv</td><td>33.6</td><td>1.82</td><td>0.68</td></tr>
 +
<tr><td>3/11 Gel Ex J23114+scFv</td><td>27.8</td><td>1.86</td><td>1.25</td></tr>
 +
<tr><td>3/11 Gel Ex pSCIC3</td><td>29.0</td><td>1.86</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Iguchi -->
 +
<div class="keln_container">
 +
<a name="0311" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/11</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Vector/(&microl)</th><th>Insert/(&microl)</th><th>Ligation High/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/11 Gel Extraction PSB1C3/ 5</td><td>3/11 Gel Extraction J23100+scfv/ 5.6</td><td>10.6</td><td>21.2</td></tr>
 +
<tr><td>3/11 Gel Extraction PSB1C3/ 5</td><td>3/11 Gel Extraction J23108+scfv/ 5</td><td>11.5</td><td>23</td></tr>
 +
<tr><td>3/11 Gel Extraction PSB1C3/ 5</td><td>3/11 Gel Extraction J23114+scfv/ 5</td><td>12.8</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0311" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/11</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>3/11 Ligation PSB1C3+J23100+scFV</td><td></td><td>DH5α /20</td><td>LB</td></tr>
 +
<tr><td>3/11 Ligation PSB1C3+J23108+scFv</td><td></td><td>DH5α /20</td><td>LB</td></tr>
 +
<tr><td>3/11 Ligation PSB1C3+J23114+scFv</td><td></td><td>DH5α /20</td><td>LB</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0307" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/7</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kbp ladder</td><td></td></tr>
 +
<tr><td>2</td><td>3/8 PCR BANcex colony1</td><td>515</td></tr>
 +
<tr><td>3</td><td>3/8 PCR BAN-cex colony2</td><td>515</td></tr>
 +
<tr><td>4</td><td>3/8 PCR BAN-clos colony1</td><td>487</td></tr>
 +
<tr><td>5</td><td>3/8 PCR BAN-clos-colony2</td><td>487</td></tr>
 +
<tr><td>6</td><td>3/8 PCR INP-cex colony 1</td><td>1405</td></tr>
 +
<tr><td>7</td><td>3/8 PCR INP-cex colony 2</td><td>1405</td></tr>
 +
<tr><td>8</td><td>3/8 PCR INP-clos colony 1</td><td>1378</td></tr>
 +
<tr><td>9</td><td>3/8 PCR INP-clos colony 2</td><td>1378</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/a/a6/T--Kyoto--3_14_No43_Electrophoresis_Yoshimoto_09_02_04_10_33.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0314" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/14</h3></span>
 +
<span class="keln_exp"><h4> PCR (Target)</h4></span>
 +
<span class="keln_researcher">Uchino, Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/25 INPcex colony2</td><td>VF 0.75    CBDcex ctl. Rv0.75</td><td>11</td><td>KAPA HiFi 12.5</td><td>25</td></tr>
 +
<tr><td>2/25 INPcex colony2</td><td>CBDcex ctl Fw 0.75  VR0.75</td><td>11</td><td>KAPA HiFi 12.5</td><td>25</td></tr>
 +
<tr><td>2/18 INPclos Gel Extraction</td><td>VF 0.75  CBD clos ctl. Rv0.75</td><td>11</td><td>KAPA HiFi 12.5</td><td>25</td></tr>
 +
<tr><td>2/18 INPclos Gel Extraction</td><td>CBDclos ctl Fw 0.75  VR0.75</td><td>11</td><td>KAPA HiFi 12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0314" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/14</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Uchino, Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>4</td><td>3/14PCR INP cex (VF CBD cex ctl.Rv)25</td><td>5</td></tr>
 +
<tr><td>5</td><td>3/14PCR INPcex (CBD cex ctl.Fw VR) 25</td><td>5</td></tr>
 +
<tr><td>6</td><td>3/14PCR INP clos(VF CBDclos ctl.Rv)25</td><td>5</td></tr>
 +
<tr><td>7</td><td>3/14PCR INPclos (CBDclos ctl.Fw VR) 25</td><td>5</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/9/98/T--Kyoto--3_14_No45_Gel_Extraction_Yoshimoto_09_02_09_48_51.1.jpg"><img src="https://static.igem.org/mediawiki/2016/5/56/T--Kyoto--3_14_No45_Gel_Extraction_Yoshimoto_09_02_09_48_51.2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0315" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/15</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/7 miniprep BAN-His-CBDcex colony2</td><td>30</td><td></td><td></td><td>1</td><td>1</td><td>4</td><td></td><td>4</td><td>40</td></tr>
 +
<tr><td>3/7miniprep BAN-His-CBDclos colony1</td><td>30</td><td></td><td></td><td>1</td><td>1</td><td>4</td><td></td><td>4</td><td>40</td></tr>
 +
<tr><td>3/7miniprep INP-His-CBDcex colony3</td><td>30</td><td></td><td></td><td>1</td><td>1</td><td>4</td><td></td><td>4</td><td>40</td></tr>
 +
<tr><td>3/'miniprep INP-His-CBDcloscolony 1</td><td>40</td><td></td><td></td><td>1</td><td>1</td><td>5</td><td></td><td>3</td><td>50</td></tr>
 +
<tr><td>2/9 miniprep J23105</td><td>61</td><td></td><td>1</td><td></td><td></td><td></td><td></td><td>0</td><td>70</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0314" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/14</h3></span>
 +
<span class="keln_exp"><h4>OE-PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/14 Gel Ex INP cex 0.5 VR ct Fw + 3/14 Gel Ex INP cex 0.5 VF ct Rv</td><td>VF1.5 VR1.5</td><td>21</td><td>25</td><td>50</td></tr>
 +
<tr><td>3/14 Gel Ex INP clos 0.5 VR ct Fw + 3/14 Gel Ex INP clos 0.5 VF ct Rv</td><td>VF1.5 VR1.5</td><td>21</td><td>25</td><td>50</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0314" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/14</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Pre Denature</td><td>95℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>78℃</td><td>20sec</td><td>35</td></tr>
 +
<tr><td>Annealing</td><td>60℃</td><td>15sec</td><td>35</td></tr>
 +
<tr><td>Extension</td><td>72℃</td><td>45sec</td><td>35</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0314" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/14</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yoshimoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/8 INP His9</td><td>VF 0.75  scFv ctl. Rv 0.75</td><td>11</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>2/8 INP His9</td><td>scFv ctl Fw 0.75 VR 0.75</td><td>11</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>2/25 INPcex colony2</td><td>VF 0.75  CBDcex ctl. Rv0.75</td><td>11</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>2/25 INPcex colony2</td><td>CBDcex ctl. Fw 0.75 VR 0.75</td><td>11</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>2/18 INPclos Gel Extraction</td><td>CBDclos ctl. Rv 0.75  VF 0.75</td><td>11</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>2/18 INPclos Gel Extraction</td><td>CBDclos ctl Fw 0.75</td><td>11</td><td>12.5</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0314" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/14</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td>5</td></tr>
 +
<tr><td>2</td><td>3/14 PCR INP His9 (VF scFv.ctl. Rv)</td><td>25</td></tr>
 +
<tr><td>3</td><td>3/14 PCR INP His9 (scFv ctl.Fw VR)</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/1/11/T--Kyoto--3_14_No173_Gel_Extraction_Yoshimoto_09_16_10_54_56%2C1.jpg"><img src="https://static.igem.org/mediawiki/2016/c/ce/T--Kyoto--3_14_No173_Gel_Extraction_Yoshimoto_09_16_10_54_56%2C2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0315" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/15</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder 5</td><td></td></tr>
 +
<tr><td>2</td><td>3/14 OE PCR CBDcex ctl 50</td><td>10</td></tr>
 +
<tr><td>3</td><td>3/14 OEPCR CBDclos ctl 50</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0315" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/15</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/25 INPcex colony2</td><td>VF /1.5</td><td>CBDcex.ctl.RV /1.5</td><td>-</td><td>22</td><td>KAPA HiFi /25</td><td>50</td></tr>
 +
<tr><td>2/25 INPcex colony2</td><td>CBDcex.ctl.Fw /1.5</td><td>VR /1.5</td><td>-</td><td>22</td><td>KAPA HiFi /25</td><td>50</td></tr>
 +
<tr><td>2/25 INPclos colony1</td><td>VF /1.5</td><td>CBDclos.ctl.Rv /1.5</td><td>-</td><td>22</td><td>KAPA HiFi /25</td><td>50</td></tr>
 +
<tr><td>2/25 INPclos colony1</td><td>CBDclos.ctl.Fw /1.5</td><td>VR /1.5</td><td>-</td><td>22</td><td>KAPA HiFi /25</td><td>50</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0315" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/15</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>4</td><td>3/15 PCR INPcex(VF 20, CBDcex ctl.Rv 20)</td><td>4</td></tr>
 +
<tr><td>5</td><td>3/15 PCR INPcex(CBDcex ctl.Fw20, VR 20)</td><td>4</td></tr>
 +
<tr><td>6</td><td>3/15 PCR INPclos(VF 20 CBDclos ctl.Rv 20)</td><td>4</td></tr>
 +
<tr><td>7</td><td>3/15 PCR INPclos(CBDclos ctl.Fw20, VR 20)</td><td>4</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0315" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/15</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Li, Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>3/11 scFv J23100 colony1,4  colony2,3(plate2)</td><td>LB(CP) 5×2</td></tr>
 +
<tr><td>3/11 scFv J23108 colony 2(plate1)  colony(plate2)</td><td>LB(CP) 5×2</td></tr>
 +
<tr><td>3/14 scFv J23114 colony 1,2,4(plate1) colony1,2,(plate2)</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Kim, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>3/15 Gelex CBDcex ctl. Fw VR</td><td>55.3</td><td>1.95</td><td>0.11</td></tr>
 +
<tr><td>3/15 Gelex CBDcex VF ctl.RV</td><td>8.2</td><td>1.74</td><td>1.04</td></tr>
 +
<tr><td>3/15Gelex CBDclos ctl Fw VR</td><td>41.2</td><td>1.85</td><td>1.51</td></tr>
 +
<tr><td>3/15Gelex CBDclos VF ctl RV</td><td>10.6</td><td>1.56</td><td>0.60</td></tr>
 +
<tr><td>3/15Gelex BANcexSP</td><td>34.9</td><td>1.70</td><td>1.84</td></tr>
 +
<tr><td>3/15Gelex BANclosSP</td><td>32.0</td><td>1.95</td><td>0.07</td></tr>
 +
<tr><td>3/15 Gelex INPcex SP</td><td>40.3</td><td>1.76</td><td>1.76</td></tr>
 +
<tr><td>3/15Gelex INPclosSP</td><td>106.8</td><td>1.83</td><td>0.19</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>3</td><td>3/15 PCR INPcex(VF 20, CBDcex ctl, RV 20)</td><td>4</td></tr>
 +
<tr><td>4</td><td>3/15 PCR INPclos(VF 20, CBDclos ctl, RV 20)</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>4</td><td>CBDcex ctl</td><td></td></tr>
 +
<tr><td>5</td><td>CBDclos ctl</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/15 Gel Ex INPcex 0.5(VF ctl.RV)  3/15 Gel Ex INPcex0.2(ctl.Fw VR)</td><td>VF1.25</td><td>VR1.25</td><td>KAPA taq 12.5</td><td>9.3</td><td></td><td>25</td></tr>
 +
<tr><td>3/15 GEl EX INPclos 0.5(VF ctl RV)  3/15Gel Ex INPclos0.2(ctl.Fw VR)</td><td>VF1.25</td><td></td><td>KAPA taq 12.5</td><td>9.3</td><td></td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/14 Gel Extraction INP-His9 VR/ct/Rv /0.5 + 3/14 Gel Extraction INP-His9 /0.5  </td><td>VF /1.5</td><td>VR /1.5</td><td>-</td><td>21</td><td>2×KAPA HiFi /25</td><td>50</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>KAPA HiFi/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/8 INP HIs9</td><td>VF 1.5 scFvctl. RV1.5</td><td>22</td><td>25</td><td>50</td></tr>
 +
<tr><td>2/8 INP His9</td><td>VR1.5 scFvctl. Fw 1.5 VR1.5</td><td>22</td><td>25</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Egashira, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>J23100 scFv c1-p1</td><td>135.7</td><td>1.85</td><td>1.76</td></tr>
 +
<tr><td>J23100 scFv C4-p1</td><td>135.1</td><td>1.86</td><td>1.85</td></tr>
 +
<tr><td>J23100 scFv c3-p2</td><td>155.0</td><td>1.87</td><td>2.04</td></tr>
 +
<tr><td>J23100 scFv c3-p2</td><td>205.9</td><td>1.76</td><td>1.32</td></tr>
 +
<tr><td>J23108 scFv c2-p1</td><td>166.5</td><td>1.88</td><td>2.06</td></tr>
 +
<tr><td>J23108 scFvc1-p2</td><td>95.7</td><td>1.84</td><td>1.83</td></tr>
 +
<tr><td>J23114 scFv c1-p1</td><td>145.7</td><td>1.80</td><td>1.99</td></tr>
 +
<tr><td>J23114 scFv c2-p1</td><td>111.7</td><td>1.90</td><td>2.16</td></tr>
 +
<tr><td>J23114 scFv c4-p1</td><td>169.2</td><td>1.90</td><td>2.11</td></tr>
 +
<tr><td>J23114 scFv c1-p2</td><td>127.4</td><td>1.87</td><td>2.00</td></tr>
 +
<tr><td>J23114 scFv c2-p2</td><td>136.2</td><td>1.90</td><td>2.13</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sukegawa -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>Sequence</h4></span>
 +
<span class="keln_researcher">Michimori</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA template/(&microl)</th><th>Primer/(&microl)</th><th>MilliQ(&microl)</th><th>Total(&microl)</th></tr>
 +
<tr><td>100A-F</td><td>3/15 miniprep J23100 scFv c1-p1 /2.2</td><td>VF /0.2</td><td>7.6</td><td>10</td></tr>
 +
<tr><td>100A-R</td><td>3/15 miniprep J23100 scFv c1-p1 /2.2</td><td>VR /0.2</td><td>7.6</td><td>10</td></tr>
 +
<tr><td>100B-F</td><td>3/15 miniprep J23100 scFv c4-p1 /2.2</td><td>VF /0.2</td><td>7.6</td><td>10</td></tr>
 +
<tr><td>100b-R</td><td>3/15 miniprep J23100 scFv c4-p1 /2.2</td><td>VR /0.2</td><td>7.6</td><td>10</td></tr>
 +
<tr><td>100C-F</td><td>3/15 miniprep J23100 scFv c2-p2 /1.9</td><td>VF /0.2</td><td>7.9</td><td>10</td></tr>
 +
<tr><td>100C-R</td><td>3/15 miniprep J23100 scFv c2-p2 /1.9</td><td>VR /0.2</td><td>7.9</td><td>10</td></tr>
 +
<tr><td>100D-F</td><td>3/15 miniprep J23100 scFv c3-p2 /1.5</td><td>VF /0.2</td><td>8.3</td><td>10</td></tr>
 +
<tr><td>100D-R</td><td>3/15 miniprep J23100 scFv c3-p2 /1.5</td><td>VR /0.2</td><td>8.3</td><td>10</td></tr>
 +
<tr><td>108A-F</td><td>3/15 miniprep J23108 scFv c2-p1 /1.8</td><td>VF /0.2</td><td>8.0</td><td>10</td></tr>
 +
<tr><td>108A-R</td><td>3/15 miniprep J23108 scFv c2-p1 /1.8</td><td>VR /0.2</td><td>8.0</td><td>10</td></tr>
 +
<tr><td>108B-F</td><td>3/15 miniprep J23108 scFv c1-p2 /3.1</td><td>VF /0.2</td><td>6.7</td><td>10</td></tr>
 +
<tr><td>108B-R</td><td>3/15 miniprep J23108 scFv c1-p2 /3.1</td><td>VR /0.2</td><td>6.7</td><td>10</td></tr>
 +
<tr><td>114A-F</td><td>3/15 miniprep J23114 scFv c1-p1 /2.1</td><td>VF /0.2</td><td>7.7</td><td>10</td></tr>
 +
<tr><td>114A-R</td><td>3/15 miniprep J23114 scFv c1-p1 /2.1</td><td>VR /0.2</td><td>7.7</td><td>10</td></tr>
 +
<tr><td>114B-F</td><td>3/15 miniprep J23114 scFv c2-p1 /2.7</td><td>VF /0.2</td><td>7.1</td><td>10</td></tr>
 +
<tr><td>114B-R</td><td>3/15 miniprep J23114 scFv c2-p1 /2.7</td><td>VR /0.2</td><td>7.1</td><td>10</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note"> 114C-R</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>Sequence</h4></span>
 +
<span class="keln_researcher">Michimori</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA template/(&microl)</th><th>Primer/(&microl)</th><th>MilliQ(&microl)</th><th>Total(&microl)</th></tr>
 +
<tr><td>114C-F</td><td>3/15 miniprep J23114 scFv c4-p1 /1.8</td><td>VF /0.2</td><td>8.0</td><td>10</td></tr>
 +
<tr><td>114C-R</td><td>3/15 miniprep J23114 scFv c4-p1 /1.8</td><td>VR /0.2</td><td>8.0</td><td>10</td></tr>
 +
<tr><td>114C-F</td><td>3/15 miniprep J23114 scFv c1-p2 /2.4</td><td>VF /0.2</td><td>7.4</td><td>10</td></tr>
 +
<tr><td>114C-R</td><td>3/15 miniprep J23114 scFv c1-p2 /2.4</td><td>VR /0.2</td><td>7.4</td><td>10</td></tr>
 +
<tr><td>114C-F</td><td>3/15 miniprep J23114 scFv c2-p2 /2.2</td><td>VF /0.2</td><td>7.6</td><td>10</td></tr>
 +
<tr><td>114C-R</td><td>3/15 miniprep J23114 scFv c2-p2 /2.2</td><td>VR /0.2</td><td>7.6</td><td>10</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note"> 114C-R</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sukegawa -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>2</td><td>3/15 PCR INP His9(VF scFv CH. Rv)</td><td>20</td></tr>
 +
<tr><td>3</td><td>3/15 PCR INP His9 (scFv ctl, Fw VR)</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder </td><td>5</td></tr>
 +
<tr><td>2</td><td>3/15 PCR INP His9 (VF  scFv ctl RV)</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sukegawa -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td></td></tr>
 +
<tr><td>2</td><td>scFv (VF ctl.Rv)</td><td></td></tr>
 +
<tr><td>3</td><td></td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Iguchi -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>OEPCR (Target)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Sample1/(&microl)</th><th>Sample2/(&microl)</th><th>Primer/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/15 Gel Extraction scfv (VF ctl.Rv)/ 0.5</td><td>3/15 Gel Extraction scfv (ctl.Fw VR)</td><td>VF/ 1.25, VR/1.25</td><td></td><td>9.3</td><td>KAPA Taq/ 12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>scFv (VF  ctl.RV)  0.5</td><td>VF 1.25  ctl.Rv) 1.25</td><td>9.5</td><td>12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0316" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/16</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/16 Gel Extraction scFv(VF.ctl.Rv) /0.25+3/16 Gel Extraction scFv(ctl.Fw.VR) /0.25</td><td>VF /1.25</td><td>VR /1.25</td><td>-</td><td>9.5</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0317" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/17</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/16 Gel Ex scFv(VF ctl.Rv)0.25, 3/15 Gel EX CBDcex(ctl.Fw VR) 0.25</td><td>VF 1.25</td><td>VR1.25</td><td>KAPA Taq 12.5</td><td>9.5</td><td></td><td>25</td></tr>
 +
<tr><td>3/16 Gel Ex scFv(VF ctl.Rv) 0.25, Gel Ex CBDclos(ctl,Fw VR) 0.25</td><td>VF 1.25</td><td>VR1.25</td><td>KAPA Taq 12.5</td><td></td><td></td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0317" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/17</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>3</td><td>3/16 OEPCR CBDcex ctl 25</td><td>500</td></tr>
 +
<tr><td>4</td><td>3/16 OEPCR CBDclos ctl 25</td><td>500</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/8/84/T--Kyoto--3_17_No59_Electrophoresis_Yoshimoto_09_03_12_05_46.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0317" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/17</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>2</td><td>CBDcex ctl 25 </td><td></td></tr>
 +
<tr><td>3</td><td>CBDclos ctl 25</td><td></td></tr>
 +
<tr><td>4</td><td>1kb ladder 5</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0317" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/17</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/16 Gel Ex scFv(VF ctl,Rv)0.1  3/15 Gel Ex CBDcex(ctl,Fw VR) 0.1  3/15 Gel Ex CBDcex(ctl,Fw VR) 0.1</td><td>VF 1.25</td><td>VR1.25</td><td>KAPA 12.5</td><td>9.8</td><td></td><td>25</td></tr>
 +
<tr><td>3/16 Gel Ex scFv(VF ctl,Rv) 0.1, 3/15 GelEx CBDclos(ctl,Fw VR) 0.1 ctl,Rv)0.1  3/15 Gel Ex CBDcex(ctl,Fw VR) 0.1</td><td>VF 1.25</td><td>VR1.25</td><td>KAPA 12.5</td><td>9.8</td><td></td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0317" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/17</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>3/8 Liquid Culture BAN-His-cex</td><td>39.3</td><td>1.91</td><td>1.67</td></tr>
 +
<tr><td>3/8 Liquid Culture BAN-His=clos</td><td>45.1</td><td>1.84</td><td>1.44</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0317" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/17</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Pre Denature</td><td>95℃</td><td>3min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>95℃</td><td>15sec</td><td>40</td></tr>
 +
<tr><td>Annealing</td><td>55℃</td><td>15sec</td><td>40</td></tr>
 +
<tr><td>Elongation</td><td>72℃</td><td>1min</td><td>40</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>1min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0317" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/17</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/17 miniprep BAN-His-cex</td><td>CBDcexFw 1.25</td><td>VR1.25</td><td>12.5</td><td>9</td><td></td><td>25</td></tr>
 +
<tr><td>3/17 miniprep BAN-His-clos</td><td>CBDclosFw 1.25</td><td>VR1.25</td><td>12.5</td><td>9</td><td></td><td>25</td></tr>
 +
<tr><td>2/9 miniprep J23105_1</td><td>VF 1.25</td><td>RFP-cex RV 1.25</td><td>12.5</td><td>9</td><td></td><td>25</td></tr>
 +
<tr><td>2/9 miniprep J23105_1</td><td>VF 1.25</td><td>RFP-clos Rv 1.25</td><td>12.5</td><td>9</td><td></td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0317" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/17</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder 5</td><td></td></tr>
 +
<tr><td>2</td><td></td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0317" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/17</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/16 Gel Extraction scFv(VF.ctl.Rv) /0.1+3/15 Gel Extraction scFv(ctl.Fw.VR) /0.1</td><td>VF /1.25</td><td>VR /1.25</td><td>-</td><td>9.8</td><td>2×KAPA HiFi /12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0318" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/18</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder </td><td>5</td></tr>
 +
<tr><td>2</td><td>3/10 PCR BAN-cex CBDcexFW VR </td><td>25</td></tr>
 +
<tr><td>3</td><td>3/10 PCR BAN-clos CBDclosFw VR </td><td>25</td></tr>
 +
<tr><td>4</td><td>3/10 PCR J23105 VF RFPcexRv </td><td>25</td></tr>
 +
<tr><td>5</td><td>3/10 PCRJ23105 VF RFPclos </td><td>25</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/6/62/T--Kyoto--3_18_No63_Gel_Extraction_Yoshimoto_09_03_12_29_48.1.jpg"><img src="https://static.igem.org/mediawiki/2016/9/98/T--Kyoto--3_18_No63_Gel_Extraction_Yoshimoto_09_03_12_29_48.2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0318" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/18</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>3/17 PCR BAN-CBDcex CBDcex.Fw+Rv</td><td>127.7</td><td>1.90</td><td>1.57</td></tr>
 +
<tr><td>3/17 PCR BAN-CBDclos CBDclos.Fw+VR</td><td>93.4</td><td>1.92</td><td>2.19</td></tr>
 +
<tr><td>3/17 PCR J23105 VF+RFP-CBDcex.Rv</td><td>104.0</td><td>1.84</td><td>1.50</td></tr>
 +
<tr><td>3/17 PCR J23105 VF+RFP-CBDclos.Rv</td><td>123.0</td><td>1.87</td><td>1.84</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0318" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/18</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Pre Denature</td><td>95℃</td><td>3min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>95℃</td><td>15sec</td><td>40</td></tr>
 +
<tr><td>Annealing</td><td>60℃</td><td>15sec</td><td>40</td></tr>
 +
<tr><td>Elongation</td><td>72℃</td><td>1min</td><td>40</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>1min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0318" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/18</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/18 Gel Extraction BAN-His-cex 0.1 cexFw+VR , 3/18Gel Extraction J23105 VF+RFP-cexRv</td><td>VF 1.25</td><td>VR 1.25</td><td>KAPATaq 12.5</td><td>9.8</td><td></td><td>25</td></tr>
 +
<tr><td>3/18 Gel Extraction BAN-His-clos 0.1 closFw+VR , 3/18Gel Extraction J23105 VF+RFP-closRv</td><td>VF 1.25</td><td>VR 1.25</td><td>KAPATaq 12.5</td><td>9.8</td><td></td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0318" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/18</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td></td></tr>
 +
<tr><td>2</td><td>3/18 OEPCR RFP-cex</td><td></td></tr>
 +
<tr><td>3</td><td>3/18 OEPCR RFP-clos</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/6/69/T--Kyoto--3_18_No66_Gel_Extraction_Yoshimoto_09_04_05_26_17.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0318" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/18</h3></span>
 +
<span class="keln_exp"><h4> PCR (Target)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/25 INP cex colony2 0.1×2</td><td>VF 0.75×2  ctl RV 0.75</td><td>10.9×2</td><td>12.5×2</td><td>25×2</td></tr>
 +
<tr><td>2/25 INP cex colony2 0.1×2</td><td>ctl Fw 0.75×2 VR 0.75×2</td><td>10.9×2</td><td>12.5×2</td><td>25×2</td></tr>
 +
<tr><td>2/25 INP cex colony2 0.1×2</td><td>VF 0.75×2 VR 0.75×2</td><td>10.9×2</td><td>12.5×2</td><td>25×2</td></tr>
 +
<tr><td>2/25 INP cex colony2 0.1×2</td><td>nothing</td><td>12.4</td><td>12.5×2</td><td>25×2</td></tr>
 +
<tr><td>2/25 INP clos colony1 0.1</td><td>VF ctl.Rv 0.75×2</td><td>10.9</td><td>12.5×2</td><td>25×2</td></tr>
 +
<tr><td>2/25 INP clos colony1 0.1</td><td>ctl.Rv 0.75×2</td><td>10.9</td><td>12.5×2</td><td>25×2</td></tr>
 +
<tr><td>2/25 INP clos colony1 0.1</td><td>VF 0.75×2 VR 0.75×2</td><td>10.9</td><td>12.5×2</td><td>25×2</td></tr>
 +
<tr><td>2/25 INP clos colony1 0.1</td><td>nothing</td><td>12.4</td><td>12.5×2</td><td>25×2</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0318" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/18</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>INP-His=CNDcex+RFP colony 1(3/16 ligation product)</td><td>VF 1.25, VR .125</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-CBDcex+RFP colony (3/16 ligation product)</td><td>VF 1.25, VR .125</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-CBDclos+RFP colony1(3/16 ligation product)</td><td>VF 1.25, VR .125</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-CBDclos+RFP colony 2(3/16 ligation product)</td><td>VF 1.25, VR .125</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-CBDcex+RFP colony1( 3/16 ligation product)</td><td>VF 1.25, VR .125</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>BAN-His-CBDcex+RFP colony2 (3/16 ligation product)</td><td>VF 1.25, VR .125</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>BAN-His-CBDclos+RFP colony1(3/16 ligation product)</td><td>VF 1.25, VR .125</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>BAN-His-CBDclos+RFP colony2(3/16 ligation product)</td><td>VF 1.25, VR .125</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0318" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/18</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Kim, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder 5</td><td></td></tr>
 +
<tr><td>2</td><td>1 kb ladder 5</td><td></td></tr>
 +
<tr><td>3</td><td>2/25 INPcex colony2 10</td><td></td></tr>
 +
<tr><td>4</td><td>2/25 INPcex colony2 10</td><td></td></tr>
 +
<tr><td>5</td><td>2/25 INPcex colony2 10</td><td></td></tr>
 +
<tr><td>6</td><td>2/25 INPcex colony210</td><td></td></tr>
 +
<tr><td>7</td><td>2/25 INPclos colony1 10</td><td></td></tr>
 +
<tr><td>8</td><td>2/25 INPclos colony1 10</td><td></td></tr>
 +
<tr><td>9</td><td>2/25 INPclos colony1 10</td><td></td></tr>
 +
<tr><td>10</td><td>2/25 INPclos colony1 10</td><td></td></tr>
 +
<tr><td>11</td><td>100bp ladder 5</td><td></td></tr>
 +
<tr><td>12</td><td>1kbp ladder 5</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/8/88/T--Kyoto--3_18_No69_Electrophoresis_Yoshimoto_09_04_05_54_18.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0318" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/18</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>INP HIs9 0.1x2</td><td>VF 0.75x2 ctl. Rv 0.75x2</td><td>10.9x2</td><td>12.5x2</td><td>25x2</td></tr>
 +
<tr><td>INP HIs9 0.1x2</td><td>ctl.Fw 1.75  VR 0.75</td><td>10.9x2</td><td>12.5x2</td><td>25x2</td></tr>
 +
<tr><td>INP HIs9 0.1x2</td><td>VF 0.75 VR 0.75</td><td>10.9x2</td><td>12.5x2</td><td>25x2</td></tr>
 +
<tr><td>INP HIs9 0.1x2</td><td>nothing</td><td>12.4</td><td>12.5x2</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0322" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/22</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /7</td><td></td></tr>
 +
<tr><td>2</td><td>INP-His-CBDcex+RFP colony1 (3/19 PCR) /20</td><td></td></tr>
 +
<tr><td>3</td><td>INP-His-CBDcex+RFP colony2 (3/19 PCR) /20</td><td></td></tr>
 +
<tr><td>4</td><td>blank</td><td></td></tr>
 +
<tr><td>5</td><td>INP-His-CBDclos+RFP colony1 (3/19 PCR) /20</td><td></td></tr>
 +
<tr><td>6</td><td>INP-His-CBDclos+RFP colony2 (3/19 PCR) /20</td><td></td></tr>
 +
<tr><td>7</td><td>blank</td><td></td></tr>
 +
<tr><td>8</td><td>BAN-His-CBDcex+RFP colony1 (3/19 PCR) /20</td><td></td></tr>
 +
<tr><td>9</td><td>BAN-His-CBDcex+RFP colony2 (3/19 PCR) /20</td><td></td></tr>
 +
<tr><td>10</td><td>blank</td><td></td></tr>
 +
<tr><td>11</td><td>BAN-His-CBDclos+RFP colony1 (3/19 PCR) /20</td><td></td></tr>
 +
<tr><td>12</td><td>BAN-His-CBDclos+RFP colony2 (3/19 PCR) /20</td><td></td></tr>
 +
<tr><td>13</td><td>blank</td><td></td></tr>
 +
<tr><td>14</td><td>100bp ladder /7</td><td></td></tr>
 +
<tr><td>15</td><td>-</td><td></td></tr>
 +
<tr><td>16</td><td>-</td><td></td></tr>
 +
<tr><td>17</td><td>-</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/7/70/T--Kyoto--3_22_No72_Electrophoresis_Notsu_10_16_12_57_19.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0322" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/22</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>7</td><td>2/25 INPcexcolony2 (5) 10</td><td></td></tr>
 +
<tr><td>8</td><td>2/25 INPcexcolony2 (6) 10</td><td></td></tr>
 +
<tr><td>9</td><td>2/25 INPcexcolony2 (7) 10</td><td></td></tr>
 +
<tr><td>10</td><td>2/25 INPcexcolony2 (8) 10</td><td></td></tr>
 +
<tr><td>11</td><td>2/25 INPclos colony1(9) 10</td><td></td></tr>
 +
<tr><td>12</td><td>2/25 INPclos colony1(10) 10</td><td></td></tr>
 +
<tr><td>13</td><td>2/25 INPclos colony1(11) 10</td><td></td></tr>
 +
<tr><td>14</td><td>2/25 INPclos colony1(12) 10</td><td></td></tr>
 +
<tr><td>16</td><td>100b ladder 5</td><td></td></tr>
 +
<tr><td>17</td><td>1kb ladder 5</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0322" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/22</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>5</td><td>3/22 PCR CBDcex(VF/ctl.Rv) 10</td><td></td></tr>
 +
<tr><td>6</td><td>3/22 PCR CBDcex(ctl.Fw/VR) 10</td><td></td></tr>
 +
<tr><td>7</td><td>3/22 PCR CBDclos(VP/ctl.RV) 10</td><td></td></tr>
 +
<tr><td>8</td><td>3/22 PCR CBDclos(ctl.Fw/VR) 10</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0322" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/22</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher"></span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/25 INPcex colony2 0.1</td><td>VF 1.5 ctl, Rv 1.5</td><td>21.9</td><td>25</td><td>50</td></tr>
 +
<tr><td>2/25 INPcex colony2 0.1</td><td>ctl.Fw 1.5 ,VR 1.5</td><td>21.9</td><td>25</td><td>50</td></tr>
 +
<tr><td>2/25 INPclos colony1 0.1</td><td>VF 1.5, ctl,Rv 1.5</td><td>21.9</td><td>25</td><td>50</td></tr>
 +
<tr><td>2/25 INPclos colony1 0.1</td><td>ctl,Fw 1.5  VR 1.5</td><td>21.9</td><td>25</td><td>50</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0322" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/22</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Notsu, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>BANcex+RFP col.1</td><td>147.8</td><td>1.89</td><td>2.18</td></tr>
 +
<tr><td>BANcex+RFP col.2 </td><td>133.9</td><td>1.87</td><td>2.18</td></tr>
 +
<tr><td>BANcex+RFP col.1</td><td>132.4</td><td>1.85</td><td>2.19</td></tr>
 +
<tr><td>BANclos+RFP col.2</td><td>126.4</td><td>1.85</td><td>2.22</td></tr>
 +
<tr><td>INPcex+RFP col.1</td><td>179.3</td><td>1.87</td><td>2.18</td></tr>
 +
<tr><td>INPcex+RFP col.2</td><td>144.9</td><td>1.87</td><td>2.19</td></tr>
 +
<tr><td>INPclos+RFPcol.1</td><td>152.2</td><td>1.87</td><td>2.23</td></tr>
 +
<tr><td>INPclos+RFPcol.2</td><td>152.4</td><td>1.86</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Koide -->
 +
<div class="keln_container">
 +
<a name="0322" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/22</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/8 INP-His9 0.1</td><td>VF 1.5</td><td>ctl.Rv 1.5</td><td>25</td><td>21.9</td><td></td><td>50</td></tr>
 +
<tr><td>2/8 INP-His9 0.1</td><td>scFv ctl.Fw 1.5</td><td>VR 1.5</td><td>25</td><td>21.9</td><td></td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0322" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/22</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Egashira, Kim</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>3/22 PCR scFv (VF/ctl.RN)</td><td>105.4</td><td>1.81</td><td>1.54</td></tr>
 +
<tr><td>3/22 PCR scFv (ctl. FV/VR)</td><td>127.4</td><td>1.87</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Iguchi -->
 +
<div class="keln_container">
 +
<a name="0322" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/22</h3></span>
 +
<span class="keln_exp"><h4>OEPCR (Target)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Sample1/(&microl)</th><th>Sample2/(&microl)</th><th>Primer/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/23 Gel Extraction scfv (VF ctl.Rv)/ 0.1</td><td>3/23 Gel Extraction scfv (ctl.Fw VR)/ 0.1</td><td>VF/ 1.25, VR/1.25</td><td></td><td>10.8</td><td>KAPA Taq/ 12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0323" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/23</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>5</td><td>3/22 PCR CBDcex(VF/ctl.Rv) 40</td><td></td></tr>
 +
<tr><td>6</td><td>3/22 PCR CBDcex(ctl.Fw/VR) 40</td><td></td></tr>
 +
<tr><td>7</td><td>3/22 PCR CBDclos(VF/ctl.Rv) 40</td><td></td></tr>
 +
<tr><td>8</td><td>3/22 PCR CBDclos(ctl.Fw/VR) 40</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0323" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/23</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Pre Denature</td><td>95℃</td><td>2min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>98℃</td><td>20sec</td><td>35</td></tr>
 +
<tr><td>Annealing</td><td>60℃</td><td>15sec</td><td>35</td></tr>
 +
<tr><td>Elongation</td><td>72℃</td><td>15sec</td><td>35</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>10℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0323" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/23</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/18 Gelex BANcex Fw+VR 0.1, 3?18 Gelex J23105 VF+cexRv 0.1</td><td>VF1.25 </td><td>VR 1.25</td><td>12.5</td><td>9.8</td><td></td><td>25</td></tr>
 +
<tr><td>3/18 Gelex BANclos closFV VR 0.1, 3/18Gelex J23105 VF+closR 0.1</td><td>VF 1.25 </td><td>VR 1.25</td><td>12.5</td><td>9.8</td><td></td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0323" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/23</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction (Measurement)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>3/22 PCR CBDcex(ctlFw/VR)</td><td>26.9</td><td>1.75</td><td>1.04</td></tr>
 +
<tr><td>3/22PCR CBDclos(ctl Fw/VR)</td><td>100.6</td><td>1.90</td><td>1.65</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0323" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/23</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>3</td><td>3/18 OEPCR BANcex cexFw+VR J23105 VF+cexRv 25</td><td></td></tr>
 +
<tr><td>4</td><td>BANclos clos Fw+VR J23105 VF+clos 25</td><td></td></tr>
 +
<tr><td>1</td><td>1kb ladder 10</td><td></td></tr>
 +
<tr><td>2</td><td>100bp ladder 10</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/1/1e/T--Kyoto--3_23_No78_Electrophoresis_Yoshimoto_09_04_06_47_22.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0323" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/23</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>4</td><td>3/23 OEPCR CBDcex ctl 25</td><td></td></tr>
 +
<tr><td>5</td><td>3/23 OEPCR CBDclos ctl 25</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0323" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/23</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/23 Gel Ex scFv(VF/ctl.Rv) 0.1, 3/23 Gel Ex CBDcex(ctl.Fw/VR) 0.1</td><td>VF 0.75 VR 0.75</td><td>10.8</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>3/23  Gel Ex scFv (VF/ctl.Rv), 3/23 Gel Ex clos(ctl.Fw/VR)</td><td>VF 0.75 VR 0.75</td><td>10.8</td><td>12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Koide -->
 +
<div class="keln_container">
 +
<a name="0323" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/23</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder 5</td><td></td></tr>
 +
<tr><td>2</td><td>100b ladder 7.5</td><td></td></tr>
 +
<tr><td>3</td><td>3/22 PCR scFv(VP/ctl.Rv) 40</td><td></td></tr>
 +
<tr><td>4</td><td>3/22 PCR scFv(ctl.Fw/VR)</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/c/c5/T--Kyoto--3_23_No199_Electrophoresis_Koide_09_19_03_50_56%2C1.jpg"><img src="https://static.igem.org/mediawiki/2016/c/c4/T--Kyoto--3_23_No199_Electrophoresis_Koide_09_19_03_50_56%2C2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0324" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/24</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Pre Denature</td><td>95℃</td><td>2min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>98℃</td><td>20sec</td><td>30</td></tr>
 +
<tr><td>Annealing</td><td>60℃</td><td>15sec</td><td>30</td></tr>
 +
<tr><td>Elongation</td><td>72℃</td><td>15sec</td><td>30</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>10℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0324" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/24</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/18 Gelcex BAN cex cexFv VR 0.2, 3/18 Gelex J23105 VF cexFv 0.2</td><td>VF1.25</td><td>VR 1.25</td><td>12.5</td><td>9.6</td><td></td><td>25</td></tr>
 +
<tr><td>3/18 Gelcex BAN clos clos Fv VR 0.2, 3/18 Gelex J23105 VF clos Rv 0.2</td><td>VF 1.25</td><td>VR 1.25</td><td>12.5</td><td>9.6</td><td></td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0324" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/24</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>3/24 OEPCR CBDcex ctl</td><td>468.6</td><td>1.82</td><td>2.06</td></tr>
 +
<tr><td>3/24 OEPCR CBDclos ctl</td><td>321.8</td><td>1.86</td><td>0.45</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0324" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/24</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>3/24 OEPCR BANcex ctl</td><td>77.2</td><td>1.81</td><td>1.40</td></tr>
 +
<tr><td>3/24 OEPCR BAN clos ctl</td><td>23.8</td><td>1.75</td><td>1.56</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0324" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/24</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>3/24 OEPCR BANcex ctl</td><td>77.2</td><td>1.81</td><td>1.40</td></tr>
 +
<tr><td>3/24 OEPCR BAN clos ctl</td><td>23.8</td><td>1.75</td><td>1.56</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0324" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/24</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>3/24 OEPCR scFv ctl</td><td>394.2</td><td>1.83</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0324" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/24</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/24 OE PCR scFv ctl </td><td>5.1</td><td>1</td><td></td><td></td><td>1</td><td>3</td><td></td><td>19.9</td><td>30</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Yoshimoto -->
 +
<div class="keln_container">
 +
<a name="0324" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/24</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>PCBTC3+scFv+ctl.</td><td></td><td>DH5α</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0325" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/25</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Egashira</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/24 OEPCR CBDcex ctl</td><td>4.3</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>20.7</td><td>30</td></tr>
 +
<tr><td>3/24 OEPCR CBDclos ctl</td><td>6.2</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>18.8</td><td>30</td></tr>
 +
<tr><td>3/15 J23100+scFv C3-P2</td><td>9.7</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>15.3</td><td>30</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0325" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/25</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/24 OEPCR BAN-CBDcex ctl</td><td>10</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>15</td><td>30</td></tr>
 +
<tr><td>3/24 OEPCR BAN-CBDclos ctl</td><td>10</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>15</td><td>30</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0325" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/25</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Vector/(&microl)</th><th>Insert/(&microl)</th><th>Ligation High/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/24 EPcut pSB1C3 /8</td><td>3/24 EPcut BAN-CBDcex.ctl /3.7</td><td>11.7</td><td>23.4</td></tr>
 +
<tr><td>3/24 EPcut pSB1C3 /7</td><td>3/24 EPcut BAN-CBDclos.ctl /4.7</td><td>11.7</td><td>23.4</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0325" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/25</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>pSB1C3+BAN-CBDcex.ctl 3/25 ligation</td><td>-</td><td>20</td><td>LB(CP)</td></tr>
 +
<tr><td>pSB1C3+BAN-CBDclos.ctl 3/25 ligation</td><td>-</td><td>20</td><td>LB(CP)</td></tr>
 +
<tr><td>pSB1C3+CBDcex 3/24 ligation</td><td>-</td><td>20</td><td>LB(CP)</td></tr>
 +
<tr><td>pSB1C3+CBDclos 3/24 ligation</td><td>-</td><td>20</td><td>LB(CP)</td></tr>
 +
<tr><td>pSB1C3+scFv.ctl 3/24 ligation</td><td>-</td><td>20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Koide -->
 +
<div class="keln_container">
 +
<a name="0328" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/28</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>BANcex ctl col.1</td><td>VF 1.25  VR 1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>BAncex ctl col.2</td><td>VF 1.25  VR 1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>BANclos ctl col.1</td><td>VF 1.25  VR 1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>BANclos ctl col.2</td><td>VF 1.25  VR 1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>CBDcex col.1</td><td>VF 1.25  VR 1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>CBDcex col.2</td><td>VF 1.25  VR 1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>CBDclos col,1</td><td>VF 1.25  VR 1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>CBDclos col.2</td><td>VF 1.25  VR 1.25</td><td>10</td><td>12.5</td><td></td></tr>
 +
<tr><td>scFv ctl col.1</td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>scFv ctl col.2</td><td></td><td></td><td></td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sukegawa -->
 +
<div class="keln_container">
 +
<a name="0328" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/28</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher"></span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder(10)</td><td></td></tr>
 +
<tr><td>2</td><td>100bb ladder(10)</td><td></td></tr>
 +
<tr><td>3</td><td>BANcex ctl col.1</td><td></td></tr>
 +
<tr><td>4</td><td>BANcex ctl col.2</td><td></td></tr>
 +
<tr><td>5</td><td>BANclos ctl col.1</td><td></td></tr>
 +
<tr><td>6</td><td>BANclos ctl col.2</td><td></td></tr>
 +
<tr><td>7</td><td>CBDcex col.1</td><td></td></tr>
 +
<tr><td>8</td><td>CBDcex col.2</td><td></td></tr>
 +
<tr><td>9</td><td>CBDclos col.1</td><td></td></tr>
 +
<tr><td>10</td><td>CBDclos col.2</td><td></td></tr>
 +
<tr><td>11</td><td>scFv ctl col.1</td><td></td></tr>
 +
<tr><td>12</td><td>scFv ctl col.2</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/2/2d/T--Kyoto--3_28_No87_Electrophoresis_Sukegawa_09_19_03_19_19.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0301" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/1</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Li, Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>100bp ladder /6</td><td></td></tr>
 +
<tr><td>2,3</td><td>J23114 /50 </td><td>193+α</td></tr>
 +
<tr><td>4,5</td><td>J23100 /50</td><td>193+α</td></tr>
 +
<tr><td>6,7</td><td>J23108 /50</td><td>193+α</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0301" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/1</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Yamamoto, Kim</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>2/29 PCR J23114</td><td>4.7</td><td>1.59</td><td>0.01</td></tr>
 +
<tr><td>2/29 PCR J23100</td><td>2.4</td><td>2.60</td><td>0.01</td></tr>
 +
<tr><td>2/29 PCR J23108</td><td>3.8</td><td>4.51</td><td>0.01</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0301" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>3/1</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>3/1 PCR purification J23100 /5</td><td>VF /0.75</td><td>J23100Rv /0.75</td><td>-</td><td>6</td><td>Prime Star Premix /12.5</td><td>25</td></tr>
 +
<tr><td>3/1 PCR purification J23108 /5</td><td>VF /0.75</td><td>J23108Rv /0.75</td><td>-</td><td>6</td><td>Prime Star Premix /12.5</td><td>25</td></tr>
 +
<tr><td>3/1 PCR purification J23114 /5</td><td>VF /0.75</td><td>J23114Rv /0.75</td><td>-</td><td>6</td><td>Prime Star Premix /12.5</td><td>25</td></tr>
 +
<tr><td>2/22 miniprep m.scFv colony3 /0.5</td><td>BAN+scFv /0.75</td><td>VR /0.75</td><td>-</td><td>10.5</td><td>Prime Star Premix /12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0722" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/22</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>7/21 transformation BclA-His-scFv colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation BclA-His-scFv colony2</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation INP-His-scFv colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation INP-His-scFv colony2</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation BclA-His-CBDcex colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation BclA-His-CBDcex colony2</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation BclA-His-CBDclos colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation BclA-His-CBDclos colony2</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation INP-His-CBDcex colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation INP-His-CBDcex colony2</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation INP-His-CBDclos colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation INP-His-CBDclos colony 2</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0725" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/25</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>7/22 liquid culture BclA-His-scFv1</td><td>183.5</td><td>1.85</td><td>2.05</td></tr>
 +
<tr><td>7/22 liquid culture BclA-His-scFv2</td><td>379.0</td><td>1.86</td><td>2.20</td></tr>
 +
<tr><td>7/22 liquid culture INP-His-scFv1</td><td>189.6</td><td>1.85</td><td>2.21</td></tr>
 +
<tr><td>7/22 liquid culture INP-His-scFv2</td><td>375.3</td><td>1.85</td><td>2.21</td></tr>
 +
<tr><td>7/22 liquid culture BclA-His-CBDcex1</td><td>91.9</td><td>1.87</td><td>1.90</td></tr>
 +
<tr><td>7/22 liquid culture BclA-His-CBDcex2</td><td>187.1</td><td>1.87</td><td>1.98</td></tr>
 +
<tr><td>7/22 liquid culture BclA-His-CBDclos1</td><td>41.2</td><td>1.88</td><td>2.21</td></tr>
 +
<tr><td>7/22 liquid culture BclA-His-CBDclos2</td><td>109.4</td><td>1.94</td><td>2.24</td></tr>
 +
<tr><td>7/22 liquid culture INP-His-CBDcex1</td><td>71.0</td><td>1.73</td><td>1.19</td></tr>
 +
<tr><td>7/22 liquid culture INP-His-CBDcex2</td><td>55.5</td><td>1.89</td><td>2.23</td></tr>
 +
<tr><td>7/22 liquid culture INP-His-CBDclos1</td><td>88.6</td><td>1.82</td><td>1.61</td></tr>
 +
<tr><td>7/22 liquid culture INP-His-CBDclos2</td><td>75.8</td><td>1.86</td><td>2.17</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0725" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/25</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/25 miniprep BclA-His-scFv1</td><td>10.9</td><td>0</td><td>0</td><td>0</td><td>0</td><td>10×L /3</td><td>0</td><td>14.1</td><td>30</td></tr>
 +
<tr><td>2/25 miniprep BclA-His-scFv2</td><td>5.3</td><td>0</td><td>1</td><td>0</td><td>1</td><td>10×H /3</td><td>3.0</td><td>16.7</td><td>30</td></tr>
 +
<tr><td>2/25 miniprep INP-His-scFv1</td><td>10.5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>10×L /3</td><td>0</td><td>14.5</td><td>30</td></tr>
 +
<tr><td>2/25 miniprep BclA-His-CBDcex2</td><td>10.7</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>14.3</td><td>30</td></tr>
 +
<tr><td>2/25 miniprep BclA-His-CBDclos2</td><td>18.3</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>6.7</td><td>30</td></tr>
 +
<tr><td>2/25 miniprep INP-His-CBDcex1</td><td>25</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>0</td><td>30</td></tr>
 +
<tr><td>2/25 miniprep INP-His-CBDclos1</td><td>22.6</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>2.4</td><td>30</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">Samples of "2/25 miniprep BclA-His-scFv1" and "2/25 miniprep INP-His-scFv1" include 2&microl of DraⅡ(Eco0109Ⅰ) (DraⅡis restriction enzyme).</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0726" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/26</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder </td><td>5</td></tr>
 +
<tr><td>2</td><td>7/25 restriction enzyme digestion BclA-His-scFv1 DraⅡcut</td><td>30</td></tr>
 +
<tr><td>3</td><td>7/25 restriction enzyme digestion BclA-His-scFv2 XPcut</td><td>30</td></tr>
 +
<tr><td>4</td><td>7/25 restriction enzyme digestion INP-His-scFv1 DraⅡcut</td><td>30</td></tr>
 +
<tr><td>5</td><td>7/25 restriction enzyme digestion BclA-His-CBDcex2 SPcut</td><td>30</td></tr>
 +
<tr><td>6</td><td>7/25 restriction enzyme digestion BclA-His-CBDclos2 SPcut</td><td>30</td></tr>
 +
<tr><td>7</td><td>7/25 restriction enzyme digestion INP-His-CBDcex1 SPcut</td><td>30</td></tr>
 +
<tr><td>8</td><td>7/25 restriction enzyme digestion INP-His-CBDclos1 SPcut</td><td>30</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/a/af/T--Kyoto--7_26_No96_Gel_Extraction_Notsu_10_02_08_39_23.1.jpg
 +
"><img src="https://static.igem.org/mediawiki/2016/1/1a/T--Kyoto--
 +
7_26_No96_Gel_Extraction_Notsu_10_02_08_39_23.2.jpg
 +
">
 
</div>
 
</div>
  
 
<!------------ Table END ------------>
 
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0726" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/26</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>7/26 Gel Extraction BclA-His-scFv2 XPcut</td><td>3.8</td><td>5.14</td><td>0.01</td></tr>
 +
<tr><td>7/26 Gel Extraction BclA-His-CBDcex2 SPcut</td><td>10.4</td><td>3.24</td><td>0.01</td></tr>
 +
<tr><td>7/26 Gel Extraction BclA-His-CBDclos2 SPcut</td><td>6.0</td><td>2.67</td><td>0.01</td></tr>
 +
<tr><td>7/26 Gel Extraction INP-His-CBDcex1 SPcut</td><td>8.1</td><td>2.03</td><td>0.02</td></tr>
 +
<tr><td>7/26 Gel Extraction INP-His-CBDclos1 SPcut</td><td>14.6</td><td>2.57</td><td>0.02</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0726" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/26</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>7/25 miniprep BclA-His-scFv1</td><td>10.9</td><td>0</td><td>0</td><td>0</td><td>0</td><td>10×L /3</td><td>0</td><td>14.1</td><td>30</td></tr>
 +
<tr><td>7/25 miniprep BclA-His-scFv1</td><td>3.6</td><td>1</td><td>0</td><td>0</td><td>1</td><td>10×H /3</td><td>0</td><td>3.4</td><td>10</td></tr>
 +
<tr><td>7/25 miniprep BclA-His-scFv1</td><td>3.6</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>6.4</td><td>10</td></tr>
 +
<tr><td>7/25 miniprep BclA-His-scFv2</td><td>5.3</td><td>0</td><td>1</td><td>0</td><td>1</td><td>10×H /3</td><td>3.0</td><td>16.7</td><td>30</td></tr>
 +
<tr><td>/25 miniprep INP-His-scFv1</td><td>10.5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>10×L /3</td><td>0</td><td>14.5</td><td>30</td></tr>
 +
<tr><td>/25 miniprep INP-His-scFv1</td><td>3.5</td><td>0</td><td>0</td><td>0</td><td>1</td><td>10×H /1</td><td>0</td><td>3.5</td><td>10</td></tr>
 +
<tr><td>/25 miniprep INP-His-scFv1</td><td>3.5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>6.5</td><td>10</td></tr>
 +
<tr><td>7/25 miniprep BclA-His-CBDcex2</td><td>10.7</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>14.3</td><td>30</td></tr>
 +
<tr><td>7/25 miniprep BclA-His-CBDclos2</td><td>18.3</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>6.7</td><td>30</td></tr>
 +
<tr><td>7/25 miniprep INP-His-CBDcex2</td><td>25</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>0</td><td>30</td></tr>
 +
<tr><td>7/25 miniprep INP-His-CBDclos2</td><td>25</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>0</td><td>30</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">Samples of "2/25 miniprep BclA-His-scFv(10.9&microl)" and "2/25 miniprep INP-His-scFv1(10.5&microl)" include 2&microl of DraⅡ(Eco0109Ⅰ) (DraⅡis restriction enzyme).</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0726" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/26</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td>5</td></tr>
 +
<tr><td>2</td><td>2/26Restriction Enzyme Digestion BclA-His-scFv1 DraⅡcut</td><td>30</td></tr>
 +
<tr><td>3</td><td>2/26Restriction Enzyme Digestion BclA-His-scFv1 XPcut</td><td>10</td></tr>
 +
<tr><td>4</td><td>2/26Restriction Enzyme Digestion BclA-His-scFv1</td><td>10</td></tr>
 +
<tr><td>5</td><td>2/26Restriction Enzyme Digestion INP-His-scFv1 DraⅡcut</td><td>30</td></tr>
 +
<tr><td>6</td><td>2/26Restriction Enzyme Digestion INP-His-scFv1EPcut</td><td>10</td></tr>
 +
<tr><td>7</td><td>2/26Restriction Enzyme Digestion INP-His-scFv1 </td><td>10</td></tr>
 +
<tr><td>-</td><td>-</td><td>-</td></tr>
 +
<tr><td>1'</td><td>1kb ladder</td><td>5</td></tr>
 +
<tr><td>2'</td><td>2/26Restriction Enzyme Digestion BclA-His-scFv2 XPcut</td><td>30</td></tr>
 +
<tr><td>3'</td><td>2/26Restriction Enzyme Digestion BclA-His-CBDcex2 SPcut</td><td>30</td></tr>
 +
<tr><td>4'</td><td>2/26Restriction Enzyme Digestion BclA-His-CBDclos2 SPcut</td><td>30</td></tr>
 +
<tr><td>5'</td><td>2/26Restriction Enzyme Digestion INP-His-CBDcex2 SPcut</td><td>30</td></tr>
 +
<tr><td>6'</td><td>2/26Restriction Enzyme Digestion INP-His-CBDclos2 SPcut</td><td>30</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/5/59/T--Kyoto--7_26_No99_Gel_Extraction_Notsu_10_02_09_03_49.1-1.jpg
 +
"><img src="https://static.igem.org/mediawiki/2016/3/36/T--Kyoto--7_26_No99_Gel_Extraction_Notsu_10_02_09_03_49.1-2.jpg"><img src="https://static.igem.org/mediawiki/2016/a/a1/T--Kyoto--7_26_No99_Gel_Extraction_Notsu_10_02_09_03_49.2-1.jpg
 +
"><img src="https://static.igem.org/mediawiki/2016/f/fe/T--Kyoto--7_26_No99_Gel_Extraction_Notsu_10_02_09_03_49.2-2.jpg
 +
">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0727" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/27</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Insert DNA(7/27purification) /&micro?</th><th>Vector DNA(7/25purification) /&micro?</th><th>Ligation High /&micro?</th><th>Total</th></tr>
 +
<tr><td>-</td><td>BclA-scFv1 DraⅡcut /7.5</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>-</td><td>INP-scFv1 DraⅡcut /7.5</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>BclAscFv2 XPcut /7.16</td><td>BclA-CBDcex2 SPcut /0.34</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>BclAscFv2 XPcut /3.75</td><td>BclA-CBDclos2 SPcut /3.75</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>BclAscFv2 XPcut /1.88</td><td>INP-CBDcex2 SPcut /5.62</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>BclAscFv2 XPcut /5.0</td><td>INP-CBDclos2 SPcut /2.5</td><td>7.5</td><td>15</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0727" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/27</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>BclAscFv1 DraⅡcut</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>INPscFv1 DraⅡcut</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>BclAscFv+BclACBDcex</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>BclAscFv+BclACBDclos</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>BclAscFv+INPCBDcex</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>BclAscfv+INPCBDclos</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0727" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/27</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>7/27 Gel Extraction BclA-His-scFv1 DraⅡcut</td><td>4.8</td><td>-7.11</td><td>1.04</td></tr>
 +
<tr><td>7/27 Gel Extraction INP-His-scFv1 DraⅡcut</td><td>40.4</td><td>1.32</td><td>1.00</td></tr>
 +
<tr><td>7/27 Gel Extraction BclA-His-scFv2 XPcut</td><td>9.3</td><td>-4.24</td><td>0.02</td></tr>
 +
<tr><td>7/27 Gel Extraction BclA-His-CBDcex2 SPcut</td><td>174.5</td><td>1.32</td><td>0.84</td></tr>
 +
<tr><td>7/27 Gel Extraction BclA-His-CBDclos2 SPcut</td><td>9.7</td><td>5.84</td><td>0.19</td></tr>
 +
<tr><td>7/27 Gel Extraction INP-His-CBDcex2 SPcut</td><td>3.7</td><td>-2.82</td><td>0.14</td></tr>
 +
<tr><td>7/27 Gel Extraction INP-His-CBDclos2 SPcut</td><td>22.6</td><td>2.51</td><td>0.21</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0728" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/28</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>DNA /&micro?</th><th>EcoR1 /&micro?</th><th>Xba1 /&micro?</th><th>Spe1 /&micro?</th><th>Pst1 /&micro?</th><th>DraⅡ /&micro?</th><th>Buffer /&micro?</th><th>BSA /&micro?</th><th>MilliQ /&micro?</th><th>Total /&micro?</th></tr>
 +
<tr><td>BclAscFv2 /5.3</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td><td>10×L /3</td><td>0</td><td>20.7</td><td>30</td></tr>
 +
<tr><td>BclAscFv2 /5.3</td><td>0</td><td>1</td><td>0</td><td>1</td><td>0</td><td>10×H /3</td><td>3</td><td>16.7</td><td>30</td></tr>
 +
<tr><td>INPscFv2 /5.3</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td><td>10×L /3</td><td>0</td><td>20.7</td><td>30</td></tr>
 +
<tr><td>BclAcex2 /10.7</td><td>0</td><td>0</td><td>1</td><td>1</td><td>0</td><td>10×M /3</td><td>0</td><td>14.3</td><td>30</td></tr>
 +
<tr><td>BclAclos1 /25</td><td>0</td><td>0</td><td>1</td><td>1</td><td>0</td><td>10×M /3</td><td>0</td><td>0</td><td>30</td></tr>
 +
<tr><td>INPcex1,2 /25</td><td>0</td><td>0</td><td>1</td><td>1</td><td>0</td><td>10×M /3</td><td>0</td><td>0</td><td>30</td></tr>
 +
<tr><td>INPclos1,2 /25</td><td>0</td><td>0</td><td>1</td><td>1</td><td>0</td><td>10×M /3</td><td>0</td><td>0</td><td>30</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0728" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/28</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Kaneko</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td></td><td>7/28 BclA-scFv2 XPcut</td><td></td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">0.5&microg/&microl<br>260/280 2.36<br>260/230 1.15</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0728" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/28</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Kaneko</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>2/22 scFv colony2 </td><td>2</td><td>DH5α /20</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0728" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/28</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Insert DNA /(&microl)                                            Vector DNA /(&microl)                                                                    Ligation high /(&microl)                                Total /(&microl)</th></tr>
 +
<tr><td>          -                                    7/28 Restriction Enzyme BclA-scFv DraⅡcut /7.5                                                7.5                                                    15</td></tr>
 +
<tr><td>          -                                    7/28 Restriction Enzyme INP-scFv DraⅡcut /7.5                                                7.5                                                    1.5</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0728" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/28</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>7/28 ligation BclA-scFv control</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>7/28 ligation INP-scFv control</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>2/22 miniprep BAN-scFv</td><td></td><td>DH5α /20</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0728" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/28</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>BclA-scFv glycerol stock </td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation INP-scFv colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation BclA-CBDcex colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation BclA-CBDclos colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation INP-CBDcex colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation INP-CBDclos colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>7/21 transformation </td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0721" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/21</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>BclA-His-scFv glycerol stock1</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-His-scFv glycerol stock2</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-His-CBDcex glycerol stock1</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-His-CBDcex glycerol stock2</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-His-CBDclos glycerol stock1</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-His-CBDclos glycerol stock2</td><td>LB(CP)</td></tr>
 +
<tr><td>INP-His-CBDcex glycerol stock1</td><td>LB(CP)</td></tr>
 +
<tr><td>INP-His-CBDcex glycerol stock2</td><td>LB(CP)</td></tr>
 +
<tr><td>INP-His-CBDclos glycerol stock1</td><td>LB(CP)</td></tr>
 +
<tr><td>INP-His-CBDclos glycerol stock2</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0721" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/21</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>2/8 miniprep INP-His-scFv</td><td>1</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>2/8 miniprep BclA-His-scFv14</td><td>1</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>2/24 miniprep BclA-His-CBDcex</td><td>1</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>2/24 miniprep BclA-His-CBDclos</td><td>1</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>2/25 miniprep INP-His-CBDcex</td><td>1</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>2/25 miniprep INP-His-CBDclos</td><td>1</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0722" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>7/22</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>2/4 parts awakening E0040</td><td>1</td><td>DH5α /20</td><td>LB(AMP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0801" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/1</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Iguchi</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>8/1 colonyPCR BclA control colony4 /10</td><td>672</td></tr>
 +
<tr><td>2</td><td>8/1 colonyPCR BclA control colony5 /10</td><td>672</td></tr>
 +
<tr><td>3</td><td>8/1 colonyPCR BclA control colony6 /10</td><td>672</td></tr>
 +
<tr><td>4</td><td>8/1 colonyPCR INP control colony4 /10</td><td>1563</td></tr>
 +
<tr><td>5</td><td>8/1 colonyPCR INP control colony5 /10</td><td>1563</td></tr>
 +
<tr><td>6</td><td>8/1 colonyPCR INP control colony6 /10</td><td></td></tr>
 +
<tr><td>7</td><td>1kb ladder /5</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/8/83/T--Kyoto--8_1_No12_Electrophoresis_Notsu_09_30_08_29_26.1.jpg"><img src="https://static.igem.org/mediawiki/2016/c/c9/T--Kyoto--8_1_No12_Electrophoresis_Notsu_09_30_08_29_26.2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0801" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/1</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Iguchi</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>8/1 miniprep BclA-scFv</td><td>8.36</td><td>0</td><td>1</td><td>0</td><td>1</td><td>10×H 3</td><td>3</td><td>13.64</td><td>30</td></tr>
 +
<tr><td>8/1 miniprep BclA-CBDcex colony1</td><td>18.2</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×H /3</td><td>0</td><td>6.8</td><td>30</td></tr>
 +
<tr><td>8/1 miniprep BclA-CBDclos colony1</td><td>21.3</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>3.7</td><td>30</td></tr>
 +
<tr><td>8/1 miniprep INP-CBDcex colony1</td><td>18.0</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>7.0</td><td>30</td></tr>
 +
<tr><td>8/1 miniprep INP-CBDclos colony1</td><td>10.8</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>16.2</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0802" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/2</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>BclA-scFv XPcut</td><td>30</td></tr>
 +
<tr><td>2</td><td>BclA-CBDcex SPcut</td><td>30</td></tr>
 +
<tr><td>3</td><td>BclA-CBDclos SPcut</td><td>30</td></tr>
 +
<tr><td>4</td><td>INP-CBDcex SPcut</td><td>30</td></tr>
 +
<tr><td>5</td><td>INP-CBDclos SPcut</td><td>30</td></tr>
 +
<tr><td>6</td><td>1kb ladder </td><td>30</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/2/28/T--Kyoto--8_2_No14_Gel_Extraction_Notsu_09_30_08_40_44.1.jpg"><img src="https://static.igem.org/mediawiki/2016/9/9f/T--Kyoto--8_2_No14_Gel_Extraction_Notsu_09_30_08_40_44.2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0802" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/2</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample                                                                            Concentration ng/&microl        260/280              260/230</th></tr>
 +
<tr><td>8/2 Electrophoresis BclA-scFv XPcut                            1.0                                      23.20                  0.12</td></tr>
 +
<tr><td>8/2 Electrophoresis BclA-CBDcex SPcut                      10.8                                    1.89                    1.35</td></tr>
 +
<tr><td>8/2 Electrophoresis BclA-CBDclos SPcut                    18.0                                    1.92                    1.88</td></tr>
 +
<tr><td>8/2 Electrophoresis INP-CBDcex SPcut                        25.4                                    1.93                    1.81</td></tr>
 +
<tr><td>8/2 Electrophoresis INP-CBDclos SPcut                      60.9</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0802" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/2</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Vector/(&microl)</th><th>Insert/(&microl)</th><th>Ligation High/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>8/2 purification BclA-CBDcex SPcut /3.5</td><td>8/2 purification BclA-scFv XPcut /4</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>8/2 purification BclA-CBDclos SPcut /3.5</td><td>8/2 purification BclA-scFv XPcut /4</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>8/2 purification INP-CBDcex SPcut /3.5</td><td>8/2 purification BclA-scFv XPcut /4</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>8/2 purification INP-CBDclos SPcut /3.5</td><td>8/2 purification BclA-scFv XPcut /4</td><td>7.5</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0802" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/2</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamamoto, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>8/2 ligation BclA-scFv+BclA-CBDcex</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>8/2 ligation BclA-scFv+BclA-CBDclos</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>8/2 ligation BclA-scFv+INP-CBDcex</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>8/2 ligation BclA-scFv+INP-CBDclos</td><td>2</td><td>DH5α /20</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0802" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/2</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>BclA-CBDcex glycerol stock</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-CBDclos glycerol stock</td><td>LB(CP)</td></tr>
 +
<tr><td>INP-CBDcex glycerol stock</td><td>LB(CP)</td></tr>
 +
<tr><td>INP-CBDclos glycerol stock</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0803" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/3</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>7/28 transformation BclA control colony5</td><td>LB(CP)</td></tr>
 +
<tr><td>7/28 transformation BclA control colony6</td><td>LB(CP)</td></tr>
 +
<tr><td>7/28 transformation INP control colony4</td><td>LB(CP)</td></tr>
 +
<tr><td>7/28 transformation INP control colony5</td><td></td></tr>
 +
<tr><td>7/28 transformation INP control colony6</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0803" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/3</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>8/1 miniprep BclA-scFv</td><td>8.36</td><td>0</td><td>1</td><td>0</td><td>1</td><td>10×H /3</td><td>3</td><td>13.64</td><td>30</td></tr>
 +
<tr><td>8/1 miniprep BclA-CBDcex</td><td>18.2</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×H /3</td><td>0</td><td>6.8</td><td>30</td></tr>
 +
<tr><td>8/1 miniprep BclA-CBDclos</td><td>21.3</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>3.7</td><td>30</td></tr>
 +
<tr><td>8/1 miniprep INP-CBDcex </td><td>18.0</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>7.0</td><td>30</td></tr>
 +
<tr><td>8/1 miniprep INP-CBDclos</td><td>10.8</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>16.2</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0803" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/3</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Vector/(&microl)</th><th>Insert/(&microl)</th><th>Ligation High/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>8/1 Restriction Enzyme Digestion BclA-CBDcex /3.5</td><td>8/1 Restriction Enzyme Digestion BclA-scFv /4</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>8/1 Restriction Enzyme Digestion BclA-CBDclos /3.5</td><td>8/1 Restriction Enzyme Digestion BclA-scFv /4</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>8/1 Restriction Enzyme Digestion INP-CBDcex /3.5</td><td>8/1 Restriction Enzyme Digestion BclA-scFv /4</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>8/1 Restriction Enzyme Digestion INP-CBDclos /3.5</td><td>8/1 Restriction Enzyme Digestion BclA-scFv /4</td><td>7.5</td><td>15</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0803" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/3</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>8/3 ligation BclA-scFv+BclA-CBDcex</td><td>15</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>8/3 ligation BclA-scFv+BclA-CBDclos</td><td>15</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>8/3 ligation BclA-scFv+INP-CBDcex</td><td>15</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>8/3 ligation BclA-scFv+INP-CBDclos</td><td>15</td><td>DH5α /20</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0804" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/4</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>8/3 transformation BclA-scFv+BclA-CBDcex colony1</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+BclA-CBDcex colony2</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+BclA-CBDcex colony3</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+BclA-CBDcex colony4</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+BclA-CBDclos colony1</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+BclA-CBDclos colony2</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+BclA-CBDclos colony3</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+BclA-CBDclos colony4</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+INP-CBDcex colony1</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+INP-CBDcex colony2</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+INP-CBDcex colony3</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+INP-CBDcex colony4</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+INP-CBDclos colony1</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+INP-CBDclos colony2</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+INP-CBDclos colony3</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>8/3 transformation BclA-scFv+INP-CBDclos colony4</td><td>VF 1.25        VR1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0804" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/4</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Predenature</td><td>95℃</td><td>1min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>95℃</td><td>15sec</td><td>30</td></tr>
 +
<tr><td>Annealing</td><td>60℃</td><td>15sec</td><td>30</td></tr>
 +
<tr><td>Extension</td><td>72℃</td><td>10sec</td><td>30</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>4℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0804" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/4</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>8/3 liquid culture BclA control colony5</td><td>137.2</td><td>1.77</td><td>1.03</td></tr>
 +
<tr><td>8/3 liquid culture BclA control colony6</td><td>117.6</td><td>1.87</td><td>1.54</td></tr>
 +
<tr><td>8/3 liquid culture INP control4</td><td>165.2</td><td>1.87</td><td>1.68</td></tr>
 +
<tr><td>8/3 liquid culture INP control5</td><td>150.6</td><td>1.90</td><td>1.81</td></tr>
 +
<tr><td>8/3 liquid culture INP control6</td><td>147.4</td><td>1.79</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0804" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/4</h3></span>
 +
<span class="keln_exp"><h4>Sequence</h4></span>
 +
<span class="keln_researcher">Yamamoto, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>No                                                                                                        primer(10&microM)          MilliQ                                  Total</th></tr>
 +
<tr><td>1 / 2                BclA_5_VF(2.18&microl) / BclA_5_VR(2.18&microl)              2&microl / 2&microl                    5.82&microl / 5.82&microl                  10&microl</td></tr>
 +
<tr><td>3 / 4                BclA_6_VF(2.55&microl) / BclA_6_VR(2.55&microl)              2&microl / 2&microl                    5.45&microl / 5.45&microl                  10&microl</td></tr>
 +
<tr><td>5 / 6                INP_4_VF(1.81&microl) / INP_4_VR(1.81&microl)                  2&microl / 2&microl                    6.19&microl / 6.19&microl                  10&microl</td></tr>
 +
<tr><td>7 / 8                INP_5_VF(1.99&microl) / INP_5_VR(1.99&microl)                  2&microl / 2&microl                    6.01&microl / 6.01&microl                  10&microl</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0804" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/4</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>8/4 colony PCR BclA-scFv+BclA-CBDcex colony1 /10</td><td>1716</td></tr>
 +
<tr><td>2</td><td>8/4 colony PCR BclA-scFv+BclA-CBDcex colony2 /10</td><td>1716</td></tr>
 +
<tr><td>3</td><td>8/4 colony PCR BclA-scFv+BclA-CBDcex colony3 /10</td><td>1716</td></tr>
 +
<tr><td>4</td><td>8/4 colony PCR BclA-scFv+BclA-CBDcex colony4 /10</td><td>1716</td></tr>
 +
<tr><td>5</td><td>8/4 colony PCR BclA-scFv+BclA-CBDclos colony1 /10</td><td>1716</td></tr>
 +
<tr><td>6</td><td>8/4 colony PCR BclA-scFv+BclA-CBDclos colony2 /10</td><td>1716</td></tr>
 +
<tr><td>7</td><td>8/4 colony PCR BclA-scFv+BclA-CBDclos colony3 /10</td><td>1716</td></tr>
 +
<tr><td>8</td><td>8/4 colony PCR BclA-scFv+BclA-CBDclos colony4 /10</td><td>1716</td></tr>
 +
<tr><td>9</td><td>100bp ladder /5</td><td>-</td></tr>
 +
<tr><td>10</td><td>1kb ladder /5</td><td>-</td></tr>
 +
<tr><td>11</td><td>-</td><td>-</td></tr>
 +
<tr><td>12</td><td>-</td><td>-</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/7/7f/T--Kyoto--8_4_No26_Electrophoresis_Notsu_10_01_11_15_31.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0804" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/4</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>8/4 colony PCR BclA-scFv+INP-CBDcex colony1 /10</td><td>2606</td></tr>
 +
<tr><td>2</td><td>8/4 colony PCR BclA-scFv+INP-CBDcex colony2 /10</td><td>2606</td></tr>
 +
<tr><td>3</td><td>8/4 colony PCR BclA-scFv+INP-CBDcex colony3 /10</td><td>2606</td></tr>
 +
<tr><td>4</td><td>8/4 colony PCR BclA-scFv+INP-CBDcex colony4 /10</td><td>2606</td></tr>
 +
<tr><td>5</td><td>8/4 colony PCR BclA-scFv+INP-CBDclos colony1 /10</td><td>2606</td></tr>
 +
<tr><td>6</td><td>8/4 colony PCR BclA-scFv+INP-CBDclos colony2 /10</td><td>2606</td></tr>
 +
<tr><td>7</td><td>8/4 colony PCR BclA-scFv+INP-CBDclos colony3 /10</td><td>2606</td></tr>
 +
<tr><td>8</td><td>8/4 colony PCR BclA-scFv+INP-CBDclos colony4 /10</td><td>2606</td></tr>
 +
<tr><td>9</td><td>100bp ladder /5</td><td>-</td></tr>
 +
<tr><td>10</td><td>1kb ladder /5</td><td></td></tr>
 +
<tr><td>11</td><td>-</td><td>-</td></tr>
 +
<tr><td>12</td><td>-</td><td>-</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/c/cc/T--Kyoto--8_4_No27_Electrophoresis_Notsu_10_01_11_18_45.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0805" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/5</h3></span>
 +
<span class="keln_exp"><h4>PartsAwaking</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Description</th><th>PartName</th><th>Backbone</th><th>Well</th><th>PlateYear</th><th>PlateNumber</th><th>MilliQ(&microl)</th></tr>
 +
<tr><td>RFP</td><td>J23105</td><td>J61002</td><td>17N</td><td>2016</td><td>4</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0805" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/5</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>8/5 Parts awakening J23105</td><td>1&microl</td><td>DH5α /20</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0809" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/9</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Iguchi, Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>8/1 miniprep BclA-scFv</td><td>8.36</td><td>0</td><td>1</td><td>0</td><td>1</td><td>10×H /3</td><td>3</td><td>13.69</td><td>30</td></tr>
 +
<tr><td>8/1 and 7/25 miniprep BclA-CBDcex</td><td>18.</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×H /3</td><td>0</td><td>6.8</td><td>30</td></tr>
 +
<tr><td>2/25 miniprep BclA-CBDclos</td><td>21.3</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>3.7</td><td>30</td></tr>
 +
<tr><td>2/25 and 7/25 minniprep INP-CBDcex</td><td>18.0</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>7.0</td><td>30</td></tr>
 +
<tr><td>8/1 miniprep INP-CBDclos</td><td>10.8</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M /3</td><td>0</td><td>16.2</td><td>30</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0809" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/9</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>8/1 transformation J23105</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0809" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/9</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>8/3 liquid culture BclA-CBDccex</td><td>225.8</td><td>1.74</td><td>1.63</td></tr>
 +
<tr><td>8/3 liquid culture BclA-CBDclos</td><td>102.3</td><td>1.80</td><td>1.66</td></tr>
 +
<tr><td>8/3 liquid culture INP-CBDcex</td><td>112.4</td><td>1.86</td><td>1.77</td></tr>
 +
<tr><td>8/3 liquid culture INP-CBDclos</td><td>42.8</td><td>1.76</td><td>0.78</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0809" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/9</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0810" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/10</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>A1</td><td>1kb ladder</td><td>10</td></tr>
 +
<tr><td>A2</td><td>BclA-scFv 8/9XPcut</td><td>30</td></tr>
 +
<tr><td>A3</td><td>BclA-CBDcex 8/9 SPcut</td><td>30</td></tr>
 +
<tr><td>A4</td><td>BclA-CBDclos 8/9 SPcu</td><td>30</td></tr>
 +
<tr><td>-</td><td>-</td><td>-</td></tr>
 +
<tr><td>B1</td><td>1kb ladder</td><td>10</td></tr>
 +
<tr><td>B2</td><td>INP-CBDcex 8/9 SPcut</td><td>30</td></tr>
 +
<tr><td>B3</td><td>INP-CBDclos 8/9 SPcut</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/b/be/T--Kyoto--8_10_No31_Gel_Extraction_Notsu_10_01_11_59_14.1.jpg"><img src="8_10_No31_Gel_Extraction_Notsu_10_01_11_59_14.2.jpg
 +
https://static.igem.org/mediawiki/2016/9/9f/T--Kyoto--">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0810" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/10</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>BclA-scFv 8/9XPcut</td><td>2.6</td><td>1.13</td><td>0.58</td></tr>
 +
<tr><td>BclA-CBDcex 8/9 SPcut</td><td>6.3</td><td>1.54</td><td>0.97</td></tr>
 +
<tr><td>BclA-CBDclos 8/9 SPcut</td><td>4.7</td><td>1.49</td><td>0.48</td></tr>
 +
<tr><td>INP-CBDcex 8/9 SPcut</td><td>4.4</td><td>1.39</td><td>0.67</td></tr>
 +
<tr><td>INP-CBDclos 8/9 SPcut</td><td>3.5</td><td>1.29</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0810" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/10</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>8/9 liquid culture J23105 (AMP) RFP colony3</td><td>178.8</td><td>1.98</td><td>1.92</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0812" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/12</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Iguchi</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>8/2 liquid culture INP-CBDclos</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-scFv glycerol stock 1</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-scFv glycerol stock 2</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-scFv glycerol stock 3</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-scFv glycerol stock 4</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0818" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/18</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Notsu</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>8/12 liquid culture INP-CBDclos</td><td>45.3</td><td>1.82</td><td>1.31</td></tr>
 +
<tr><td>8/12 liquid culture BclA-scFv glycerol stock 1</td><td>101.6</td><td>1.59</td><td>0.71</td></tr>
 +
<tr><td>8/12 liquid culture BclA-scFv glycerol stock 2</td><td>100.7</td><td>1.67</td><td>0.78</td></tr>
 +
<tr><td>8/12 liquid culture BclA-scFv glycerol stock 3</td><td>77.8</td><td>1.60</td><td>0.70</td></tr>
 +
<tr><td>8/12 liquid culture BclA-scFv glycerol stock 4</td><td>41.0</td><td>1.87</td><td>1.27</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0820" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/20</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Ohara, Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>BclA-CBDcex</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-CBDclos</td><td>LB(CP)</td></tr>
 +
<tr><td>INP-CBDcex</td><td>LB(CP)</td></tr>
 +
<tr><td>INP-CBDclos</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0821" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/21</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>BclA-scFv ①(glisto）</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-scFv ②</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-scFv ③</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-scFv  ④</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0822" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/22</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>8/22 Restraction Ensyme Digestion 1.1kb ladder</td><td>5</td><td></td></tr>
 +
<tr><td>8/22 Restraction Ensyme Digestion Bcla-cex</td><td>25</td><td>2520</td></tr>
 +
<tr><td>8/22 Restraction Ensyme Digestion Bcla-clos</td><td>25</td><td>2520</td></tr>
 +
<tr><td>8/22 Restraction Ensyme Digestion INP-cex(s,pcut)</td><td>25</td><td>3500</td></tr>
 +
<tr><td>8/22 Restraction Ensyme Digestion INP-clos</td><td>25</td><td>3500</td></tr>
 +
<tr><td>8/22 Restraction Ensyme Digestion INP-cex</td><td>25</td><td>2541</td></tr>
 +
<tr><td>8/22 Restraction Ensyme Digestion J23105 (AMP) RFP 8/21 old</td><td>25</td><td>950</td></tr>
 +
<tr><td>8/22 Restraction Ensyme Digestion Liquid culture Bcla-scfv glisto 4</td><td>25</td><td>932</td></tr>
 +
<tr><td>8/22 Restraction Ensyme Digestion Bcla-scfv glisto 2</td><td>25</td><td>932</td></tr>
 +
<tr><td>100ladder</td><td>5</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/9/9f/T--Kyoto--8_22_No49_Electrophoresis_Sassa_08_31_11_29_24.1.jpg"><img src="https://static.igem.org/mediawiki/2016/a/a4/T--Kyoto--8_22_No49_Electrophoresis_Sassa_08_31_11_29_24.2.jpg"><img src="https://static.igem.org/mediawiki/2016/f/fa/T--Kyoto--8_22_No49_Electrophoresis_Sassa_08_31_11_29_24.3.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0822" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/22</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>8/12 Liquid cultureBclA-scFv glisto ④</td><td>38</td><td>0</td><td>1</td><td>0</td><td>1</td><td>5</td><td>5</td><td></td><td>50</td></tr>
 +
<tr><td>8/12 Liquid cultureBclA-scFv glisto  ②</td><td>38</td><td>0</td><td>1</td><td>0</td><td>1</td><td>5</td><td>5</td><td>0</td><td>50</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0822" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/22</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yoshimoto, Ohara, Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>BclA control</td><td>LB(CP)</td></tr>
 +
<tr><td>INP control</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0822" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/22</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>2/9 J23105 RFP</td><td></td><td>DH5α /20</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0822" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/22</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Yoshimoto</span>
 +
<table class="keln_table">
 +
<tr><th>1st</th><th></th><th></th><th></th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>BclAclos+</td><td>0.208/0.206/0.206</td><td>0.131/0.135/0.14</td><td>0.111/0.112/0.109</td></tr>
 +
<tr><td>INPclos+</td><td>0.183/0.186/0.184</td><td>0.239/0.23/0.22</td><td>0.158/0.155/0.169</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0822" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/22</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Yoshimoto</span>
 +
<table class="keln_table">
 +
<tr><th>2nd</th><th></th><th></th><th></th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>BclAclos+</td><td>0.208/0.205/0.204</td><td>0.078/0.074/0.075</td><td>0.056/0.056/0.039</td></tr>
 +
<tr><td>INPclos+</td><td>0.183/0.181/0.187</td><td>0.093/0.097/0.099</td><td>0.067/0.073/0.069</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0823" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/23</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>2nd</th><th></th><th></th><th></th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>BclAcex+ (15min)</td><td>0.247/0.247/0.248</td><td>0.221/0.224/0.218</td><td>0.085/0.075/0.073</td></tr>
 +
<tr><td>BclAcex+ (30min)</td><td>0.247/0.247/0.248</td><td>0.243/0.229/0.228</td><td>0.034/0.048/0.038</td></tr>
 +
<tr><td>BclAcex+ (60min)</td><td>0.247/0.247/0.248</td><td>0.224/0.221/0.217</td><td>0.065/0.064/0.065</td></tr>
 +
<tr><td>BclAcex- (15min)</td><td>0.247/0.247/0.248</td><td>0.257/0.253/0.258</td><td></td></tr>
 +
<tr><td>BclAcex- (30min)</td><td>0.247/0.247/0.248</td><td>0.236/0.230/0.235</td><td></td></tr>
 +
<tr><td>BclAcex- (60min)</td><td>0.247/0.247/0.248</td><td>0.249/0.248/0.247</td><td></td></tr>
 +
<tr><td> cellulose only (60min)</td><td></td><td>0.021/0.021/0.023</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0823" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/23</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>4th</th><th></th><th></th><th></th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>BclAcex+ (2ml tube)</td><td>0.227/0.226/0.225</td><td>0.108/0.108/0.108</td><td>0.076/0.074/0.081</td></tr>
 +
<tr><td>BclActl+ (2ml tube)</td><td>0.186/0.203/0.189</td><td>0.082/0.081/0.082</td><td>0.055/0.046/0.044</td></tr>
 +
<tr><td>BclAcex- (2ml tube)</td><td>0.235/0.237/0.233</td><td>0.261/0.262/0.254</td><td></td></tr>
 +
<tr><td>BclAcex+ (1.5ml tube)</td><td>0.237/0.226/0.236</td><td>0.159/0.157/0.158</td><td>0.066/0.063/0.065</td></tr>
 +
<tr><td>BclActl+ (1.5ml tube)</td><td>0.177/0.173/0.179</td><td>0.112/0.109/0.113</td><td>0.080/0.076/0.095</td></tr>
 +
<tr><td>BclAcex- (1.5ml tube)</td><td>0.240/0.235/0.238</td><td>0.255/0.254/0.253</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0824" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/24</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>8/5 transormation J2305</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0824" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/24</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>8/24 BclA-scFv 1</td><td>LB(CP)</td></tr>
 +
<tr><td>8/24 BclA-scFv 2</td><td>LB(CP)</td></tr>
 +
<tr><td>8/24 BclA-scFv 3</td><td>LB(CP)</td></tr>
 +
<tr><td>8/24 BclA-scFv 4</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0824" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/29</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>1.  8/23Liquidculture Bcla scFv</td><td>111.6</td><td>1.98</td><td>1.82</td></tr>
 +
<tr><td> 2. 8/23Liquidculture Bcla scFv  </td><td>143.1</td><td>2.00</td><td>2.03</td></tr>
 +
<tr><td> 2'. 8/23Liquidculture Bcla scFv</td><td>141.6</td><td>2.04</td><td>2.10</td></tr>
 +
<tr><td> 3. 8/23Liquidculture Bcla scFv</td><td>145.2</td><td>2.07</td><td>2.01</td></tr>
 +
<tr><td>8/23Liquidculture J23105</td><td>63.1</td><td>2.01</td><td>1.69</td></tr>
 +
<tr><td>8/23Liquidculture J23105</td><td>68.1</td><td>2.00</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0824" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/24</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>5th</th><th></th><th></th><th></th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>INPcex+</td><td>0.195/0.191/0.192</td><td>0.103/0.102/0.109</td><td>0.059/0.051/0.055</td></tr>
 +
<tr><td>INPcex-</td><td>0.190/0.191/0.205</td><td>0.186/0.187/0.186</td><td></td></tr>
 +
<tr><td>INPclos+</td><td>0.187/0.187/0.188</td><td>0.121/0.122/0.126</td><td>0.051/0.058/0.059</td></tr>
 +
<tr><td>INPclos-</td><td>0.196/0.195/0.191</td><td>0.206/0.204/0.202</td><td></td></tr>
 +
<tr><td>INPctl+</td><td>0.172/0.163/0.162</td><td>0.116/0.177/0.124</td><td>0.066/0.063/0.062</td></tr>
 +
<tr><td>INPctl-</td><td>0.171/0.175/0.165</td><td>0.167/0.182/0.171</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0824" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/24</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>6th</th><th>null</th><th>null</th><th>null</th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>INPcex+</td><td>0.184/0.184/0.189</td><td>0.104/0.106/0.106</td><td>0.044/0.044/0.056</td></tr>
 +
<tr><td>INPcex-</td><td>0.186/0.190/0.184</td><td>0.182/0.181/0.184</td><td></td></tr>
 +
<tr><td>INPclos+</td><td>0.184/0.189/0.185</td><td>0.097/0.098/0.099</td><td>0.037/0.037/0.038</td></tr>
 +
<tr><td>INPclos-</td><td>0.187/0.192/0.190</td><td>0.185/0.184/0.189</td><td></td></tr>
 +
<tr><td>INPctl+</td><td>0.173/0.184/0.180</td><td>0.095/0.099/0.095</td><td>0.045/0.046/0.046</td></tr>
 +
<tr><td>INPctl-</td><td>0.185/0.184/0.183</td><td>0.175/0.175/0.175</td><td></td></tr>
 +
<tr><td>BclAcex+</td><td>0.170/0.168/0.174</td><td>0.099/0.094/0.092</td><td>0.037/0.032/0.036</td></tr>
 +
<tr><td>BclAcex-</td><td>0.186/0.189/0.171</td><td>0.164/0.167/0.164</td><td></td></tr>
 +
<tr><td>cellulose only</td><td>0.006/0.005/0.006</td><td>0.013/0.010/0.006</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0825" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/25</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>7th</th><th>null</th><th>null</th><th>null</th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>BclAcex+</td><td>0.181/0.181/0.181</td><td>0.192/0.196/0.195</td><td>0/0/0</td></tr>
 +
<tr><td>BclAcex-</td><td>0.149/0.150/0.151</td><td>0.221/0.213/0.211</td><td></td></tr>
 +
<tr><td>BclActl+</td><td>0.169/0.165/0.167</td><td>0.154/0.156/0.157</td><td>0.012/0.013/0.011</td></tr>
 +
<tr><td>BclActl-</td><td>0.151/0.153/0.154</td><td>0.187/0.178/0.173</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0826" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/26</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>8/10 miniprep J23105 /3</td><td>VF /1.25</td><td>VR /1.25</td><td>-</td><td>7</td><td>KAPA2GFAST /12.5</td><td>25</td></tr>
 +
<tr><td>8/24 OD0.206 INP-CBDclos /3</td><td>VF /1.25</td><td>VR /1.25</td><td>-</td><td>7</td><td>KAPA2GFAST /12.5</td><td>25</td></tr>
 +
<tr><td>8/24 OD0.202 INP-CBDcex</td><td>VF /1.25</td><td>VR /1.25</td><td>-</td><td>7</td><td>KAPA2GFAST /12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0826" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/26</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Yamamoto, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder /5</td><td></td></tr>
 +
<tr><td>2</td><td>8/26 PCR INP-CBDcex VFVR /15</td><td></td></tr>
 +
<tr><td>3</td><td>8/26 PCR INP-CBDclos VFVR /15</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/4/4b/T--Kyoto--8_26_No62_Electrophoresis_Notsu_10_01_02_14_38.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0826" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/26</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>8/26 Gel Extraction Bcla cex</td><td>8.3</td><td>1.52</td><td>0.25</td></tr>
 +
<tr><td>8/26 Gel Extraction Bcla clos</td><td>10.2</td><td>1.70</td><td>0.26</td></tr>
 +
<tr><td>8/26 Gel Extraction INP cex</td><td>11.4</td><td>1.76</td><td>0.15</td></tr>
 +
<tr><td>8/26 Gel Extraction INP clos</td><td>7.7</td><td>1.62</td><td>0.09</td></tr>
 +
<tr><td>8/26 Gel Extraction Bcla scFv glisto</td><td>7.0</td><td>1.59</td><td>0.20</td></tr>
 +
<tr><td>8/26 Gel Extraction Bcla scFv glisto</td><td>14.1</td><td>6.33</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0826" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/26</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Yamamoto, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>8/26 Gel Extraction Bcla cex</td><td>8.3</td><td>1.52</td><td>0.25</td></tr>
 +
<tr><td>8/26 Gel Extraction Bcla clos</td><td>10.2</td><td>1.70</td><td>0.26</td></tr>
 +
<tr><td>8/26 Gel Extraction INP cex</td><td>11.4</td><td>1.76</td><td>0.15</td></tr>
 +
<tr><td>8/26 Gel Extraction  INP clos</td><td>7.7</td><td>1.62</td><td>0.09</td></tr>
 +
<tr><td>8/26 Gel Extraction Bcla scFv glisto</td><td>7.0</td><td>1.59</td><td>0.20</td></tr>
 +
<tr><td>8/26 Gel Extraction Bcla scFv glisto</td><td>14.1</td><td>6.33</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0826" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/26</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Yoshimoto</span>
 +
<table class="keln_table">
 +
<tr><th>8th</th><th>null</th><th>null</th><th>null</th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>BclAcex+</td><td>0.183/0.185/0.188</td><td>0.116/0.115/0.114</td><td>0.086/0.086/0.089</td></tr>
 +
<tr><td>BclActl+</td><td>0.116/0.115/0.114</td><td>0.096/0.092/0.087</td><td>0.09/0.079/0.084</td></tr>
 +
<tr><td>cellulose only</td><td></td><td>0.037/0.035/0.037</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0826" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/26</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>9th</th><th>null</th><th>null</th><th>null</th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>BclAcex+</td><td>0.186/0.186/0.184</td><td>0.170/0.169/0.170</td><td>0.021/0.024/0.020</td></tr>
 +
<tr><td>BclAcex-</td><td>0.154/0.149/0.149</td><td>0.142/0.143/0.141</td><td></td></tr>
 +
<tr><td>BclActl+</td><td>0.194/0.189/0.189</td><td>0.167/0.164/0.159</td><td>0.010/0.009/0.008</td></tr>
 +
<tr><td>BclActl-</td><td>0.162/0.168/0.169</td><td>0.159/0.162/0.165</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0829" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/29</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>Insert DNA</th><th>vector DNA</th><th>Ligation High</th><th>Total</th></tr>
 +
<tr><td>8/26 Gel Extraction BclAscFv /3.4</td><td>8/26 Gel Extraction Bcla CBDcex /4.1</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>8/26 Gel Extraction BclAscFv /3.1</td><td>8/26 Gel Extraction Bcla CBD clos /4.4</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>8/26 Gel Extraction BclAscFvBclAscFv /3.8</td><td>8/26 Gel Extraction Bcla CBD cex /4.7</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>8/26 Gel Extraction BclAscFv /2.6</td><td>8/26 Gel Extraction Bcla CBD clos /4.9</td><td>7.5</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0829" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/29</h3></span>
 +
<span class="keln_exp"><h4>Western Blotting</h4></span>
 +
<span class="keln_researcher">Ohara, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>lane</th><th>sample</th></tr>
 +
<tr><td>1</td><td>Marker</td></tr>
 +
<tr><td>2</td><td>Sano VLP 0.5&microl</td></tr>
 +
<tr><td>3</td><td>Sano VLP 0.2&microl</td></tr>
 +
<tr><td>4</td><td>Sano VLP 0.1&microl</td></tr>
 +
<tr><td>5</td><td>Sano VLP 0.05&microl</td></tr>
 +
<tr><td>6</td><td>Sano VLP 0.02&microl</td></tr>
 +
<tr><td>7</td><td>Sano VLP 0.05&microl</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/0/0e/T--Kyoto--8_29_No68_General_Miyazaki_10_17_05_42_18.jpg">
 +
</div>
 +
<p class="keln_note">blotting CC<br>1'antibody time/buffer  1:3000 6ml 1h<br>2'antibody time/buffer  1:10000 10ml 1h<br>Blocking 1day<br>transfer 100mA  1h</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0829" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/29</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>BclA-CBDcex</td><td>VF /1.25</td><td>VR /1.25</td><td>-</td><td>10</td><td>KAPA2GFAST /12.5</td><td>25</td></tr>
 +
<tr><td>BclA-CBDclos</td><td>VF /1.25</td><td>VR /1.25</td><td>-</td><td>10</td><td>KAPA2GFAST /12.5</td><td>25</td></tr>
 +
<tr><td>INP-CBDcex</td><td>VF /1.25</td><td>VR /1.25</td><td>-</td><td>10</td><td>KAPA2GFAST /12.5</td><td>25</td></tr>
 +
<tr><td>INP-CBDclos</td><td>VF /1.25</td><td>VR /1.25</td><td>-</td><td>10</td><td>KAPA2GFAST /12.5</td><td>25</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/8/85/T--Kyoto--8_29_No69_PCR_%28Target%29_Notsu_10_01_02_11_53.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0829" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/29</h3></span>
 +
<span class="keln_exp"><h4>ligation</h4></span>
 +
<span class="keln_researcher">Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>Insert DNA</th><th>Vector DNA</th><th>Ligation High </th><th>Total</th></tr>
 +
<tr><td>BclAscFv XPcut  6.3</td><td>INPCBDcex SPcut  1.2</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>BclAscFv XPcut  6.4</td><td>INPCBDclos SPcut  1.1</td><td>7.5</td><td>15</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0829" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/29</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Sassa, Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>BclA scFv 1</td><td>8.4</td><td>1.63</td><td>0.11</td></tr>
 +
<tr><td>BclA scFv 2</td><td>5.6</td><td>1.91</td><td>0.04</td></tr>
 +
<tr><td>INP CBDcex</td><td>44.8</td><td>1.90</td><td>0.70</td></tr>
 +
<tr><td>INP CBDclos</td><td>33.1</td><td>1.82</td><td>0.68</td></tr>
 +
<tr><td>BclA cex</td><td>26.3</td><td>1.93</td><td>0.62</td></tr>
 +
<tr><td>BclA clos </td><td>24.4</td><td>1.93</td><td>0.66</td></tr>
 +
<tr><td>J23105 VF VR</td><td>33.2</td><td>1.88</td><td>0.24</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0801" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/1</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>7/28 transformaton BclA control colony4</td><td>VF/1.25  VR/1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>7/28 transformaton BclA control colony5</td><td>VF/1.25  VR/1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>7/28 transformaton BclA control colony6</td><td>VF/1.25  VR/1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>7/28 transformaton INP control colony4</td><td>VF/1.25  VR/1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>7/28 transformaton INP control colony5</td><td>VF/1.25  VR/1.25</td><td>10</td><td>12.5</td><td>25</td></tr>
 +
<tr><td>7/28 transformaton INP control colony6</td><td>VF/1.25  VR/1.25</td><td>10</td><td>12.5</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0801" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/1</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Predenature</td><td>95℃</td><td>1min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>95℃</td><td>15sec</td><td>30</td></tr>
 +
<tr><td>Annealing</td><td>60℃</td><td>15sec</td><td>30</td></tr>
 +
<tr><td>Extension</td><td>72℃</td><td>10sec</td><td>30</td></tr>
 +
<tr><td>Final Elogation</td><td>72℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td></td><td>-</td><td>1</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0801" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>8/1</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>7/28 liquid culture BclA-scFv</td><td>239.1</td><td>1.87</td><td>1.78</td></tr>
 +
<tr><td>7/28 liquid culture INP-scFv colony1</td><td>263.4</td><td>1.86</td><td>1.99</td></tr>
 +
<tr><td>7/28 liquid culture E0040 colony1</td><td>98.8</td><td>1.84</td><td>2.15</td></tr>
 +
<tr><td>7/28 liquid culture BclA-CBDcex colony1</td><td>109.9</td><td>1.82</td><td>1.37</td></tr>
 +
<tr><td>7/28 liquid culture BclA-CBDclos colony1</td><td>94.1</td><td>1.89</td><td>1.93</td></tr>
 +
<tr><td>7/28 liquid culture INP-CBDcex colony1</td><td>111.4</td><td>1.82</td><td>1.39</td></tr>
 +
<tr><td>7/28 liquid culture INP-CBDclos colony1</td><td>185.1</td><td>1.81</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0905" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/5</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>BclA-CBDcex /20ml</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-control /20ml</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0905" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/5</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Sassa, Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>9/5 electrophoresis 2 </td><td>J23105 RFP XPcut backbone</td><td></td></tr>
 +
<tr><td>9/5 electrophoresis 2</td><td>J23105 RFP XPcut insert</td><td></td></tr>
 +
<tr><td>9/5 electrophoresis 1</td><td>BclAcex SPcut</td><td></td></tr>
 +
<tr><td>9/5 electrophoresis 2</td><td>BclAclos SPcut</td><td></td></tr>
 +
<tr><td>9/5 electrophoresis 3</td><td>INPcex SPcut</td><td></td></tr>
 +
<tr><td>9/5 electrophoresis 4</td><td>INPclos SPcut</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0906" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/6</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>Bclascfv+BclaCBDcex colony1</td><td>vf1.25 vr1.25</td><td>10</td><td>KAPA2GFAST 12.5</td><td>25</td></tr>
 +
<tr><td>〃colony2</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony3</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony4</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>Bclascfv+BclaCBDclos colony1</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony2</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony3</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony4</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>Bclascfv+INPCBDcex colony1</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony2</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony3</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony4</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>Bclascfv+INPCBDclos colony1</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony2</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony3</td><td>〃</td><td>〃</td><td>〃</td><td>onaji  </td></tr>
 +
<tr><td>〃colony4</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0906" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/6</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Insert DNA(&microl)</th><th>Vector DNA/(&microl)</th><th>Ligation High/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>9/5 Gel Extraction BclA cex SPcut/7</td><td>9/5 Gel Extraction J23105 xpcut 36/0.5</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>9/5 Gel Extraction BclA clos SPcut/7</td><td>9/5 Gel Extraction J23105 xpcut 36/0.5</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>9/5 Gel Extraction INP cex  SPcut/7</td><td>9/5 Gel Extraction J23105 xpcut 36/0.5</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>9/5 Gel Extraction INP clos  SPcut/7</td><td>9/5 Gel Extraction J23105 xpcut 36/0.5</td><td>7.5</td><td>15</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0906" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/6</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher"></span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder/5</td><td>0</td></tr>
 +
<tr><td>2</td><td>9/6 PCR scfv+Bclacex colony1/15</td><td>0</td></tr>
 +
<tr><td>3</td><td>〃2/15</td><td>0</td></tr>
 +
<tr><td>4</td><td>〃3/15</td><td>0</td></tr>
 +
<tr><td>5</td><td>〃4/15</td><td>0</td></tr>
 +
<tr><td>6</td><td>9/6 PCR scfv+Bclaclos colony1</td><td>0</td></tr>
 +
<tr><td>7</td><td>〃2</td><td>0</td></tr>
 +
<tr><td>8</td><td>〃3</td><td>0</td></tr>
 +
<tr><td>9</td><td>〃4</td><td>0</td></tr>
 +
<tr><td>10</td><td>9/6 PCR scfv+INPcex colony1</td><td>0</td></tr>
 +
<tr><td>11</td><td>〃2</td><td>0</td></tr>
 +
<tr><td>12</td><td>〃3</td><td>0</td></tr>
 +
<tr><td>13</td><td>〃4</td><td>0</td></tr>
 +
<tr><td>14</td><td>9/6 PCR scfv+INPclos colony1</td><td>0</td></tr>
 +
<tr><td>15</td><td>〃2</td><td>0</td></tr>
 +
<tr><td>16</td><td>〃3</td><td>0</td></tr>
 +
<tr><td>17</td><td>〃4</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/a/a4/T--Kyoto--9_6_No81_Electrophoresis_Kosaka_09_30_11_22_50.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0906" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/6</h3></span>
 +
<span class="keln_exp"><h4>Western Blotting</h4></span>
 +
<span class="keln_researcher">Ohara, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>lane</th><th>sample</th></tr>
 +
<tr><td>1</td><td>Marker</td></tr>
 +
<tr><td>2</td><td>Sano VLP 0.5&microl</td></tr>
 +
<tr><td>3</td><td>Sano VLP 0.2&microl</td></tr>
 +
<tr><td>4</td><td>Sano VLP 0.1&microl</td></tr>
 +
<tr><td>5</td><td>Sano VLP 0.05&microl</td></tr>
 +
<tr><td>6</td><td>Sano VLP 0.02&microl</td></tr>
 +
<tr><td>7</td><td>Sano VLP 0.05&microl</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/3/33/T--Kyoto--9_6_No82_General_Miyazaki_10_17_05_43_53.jpg">
 +
</div>
 +
<p class="keln_note">blotting CC<br>1'antibody time/buffer  1:3000 6ml 1h<br>2'antibody time/buffer  1:10000 10ml 1h<br>Blocking 1day<br>transfer 100mA  1h</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0906" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/6</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>10th</th><th>null</th><th>null</th><th>null</th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>BclAcex+</td><td>0.239</td><td>0.222</td><td></td></tr>
 +
<tr><td>BclActl+</td><td>0.212</td><td>0.190</td><td></td></tr>
 +
<tr><td>BclAcex-</td><td>0.239</td><td>0.233</td><td></td></tr>
 +
<tr><td>BclActl-</td><td>0.221</td><td>0.210</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0906" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/6</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>11th</th><th>null</th><th>null</th><th>null</th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>BclAcex+ (NaCl)</td><td>0.202</td><td>0.176</td><td></td></tr>
 +
<tr><td>BclAcex- (NaCl)</td><td>0.202</td><td>0.206</td><td></td></tr>
 +
<tr><td>BclAcex- (NaCl)</td><td>0.202</td><td>0.264</td><td></td></tr>
 +
<tr><td>BclAcex+ (PBS buffer)</td><td>0.212</td><td>0.179</td><td></td></tr>
 +
<tr><td>BclAcex- (PBS buffer)</td><td>0.212</td><td>0.209</td><td></td></tr>
 +
<tr><td>BclActl+ (PBS buffer)</td><td>0.211</td><td>0.172</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0907" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/7</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>Bclascfv-BclaCBDcex colony5</td><td>vf12.5 vr12.5</td><td>10</td><td>KAPA2GFASTHSRM 12.5</td><td>25</td></tr>
 +
<tr><td>〃6</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃7</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃8</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃9</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃10</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>9/5 Bclascfv-BclaCBDclos 5</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃6</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>9/5 Bclascfv-INPCBDcex1</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃2</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃3</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃4</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>Bclascfv-INPCBDclos1</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃2</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃3</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃4</td><td>=</td><td>=</td><td>=</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/c/c6/T--Kyoto--9_7_No83_Colony_PCR_%28typeA%29_Kosaka_09_30_11_33_11.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0907” class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/7</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>Bcla-CBDcex</td><td>99.7</td><td>1.94</td><td>2.08</td></tr>
 +
<tr><td>Bcla-CBDclos</td><td>73.5</td><td>1.96</td><td>2.15</td></tr>
 +
<tr><td>INP-CBDcex</td><td>45.3</td><td>1.99</td><td>2.14</td></tr>
 +
<tr><td>INP-CBDclos</td><td>49.6</td><td>1.89</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0907" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/7</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>8/5 miniprep INP-CBDcex</td><td>25</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M/3</td><td>0</td><td>0</td><td>30</td></tr>
 +
<tr><td>8/5 miniprep INP-CBDclos</td><td>25</td><td>0</td><td>0</td><td>1</td><td>1</td><td>10×M/3</td><td>0</td><td>0</td><td>30</td></tr>
 +
<tr><td>9/13 miniprep BclA-scFv1</td><td>22</td><td>0</td><td>1</td><td>0</td><td>1</td><td>10×H/3</td><td>3</td><td>0</td><td>30</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0907" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/7</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>9/5 Transformation BclascFv-BclaCBDcex colony5</td><td>LB(CP)/5ml</td></tr>
 +
<tr><td>〃BclascFv-BclaCBDclos colony4</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0907" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/7</h3></span>
 +
<span class="keln_exp"><h4>SEM</h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>number</th><th>sample</th><th>OD600</th><th>VLP(&microl)</th></tr>
 +
<tr><td>1</td><td>BclAscFv 3c5</td><td>0.990</td><td>1</td></tr>
 +
<tr><td>2</td><td>BclAscFv 1c5</td><td>1.001</td><td>1</td></tr>
 +
<tr><td>3</td><td>INPscFv 3c5</td><td>1.062</td><td>1</td></tr>
 +
<tr><td>4</td><td>BclA CTL</td><td>1.053</td><td>1</td></tr>
 +
<tr><td>5</td><td>INP CTL</td><td>0.952</td><td>1</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0907" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/7</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>Bcla-CBDcex</td><td>LB(CP)/30ml</td></tr>
 +
<tr><td>Bcla-control</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0907" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/7</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>12th</th><th>null</th><th>null</th><th>null</th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>BclAcex+ (PBS buffer)</td><td>0.195/0.192</td><td>0.188/0.189</td><td></td></tr>
 +
<tr><td>BclActl+ (PBS buffer)</td><td>0.198/0.195</td><td>0.192/0.189</td><td></td></tr>
 +
<tr><td>BclAcex+ (CB buffer)</td><td>0.186/0.184</td><td>0.176/0.173</td><td></td></tr>
 +
<tr><td>BclActl+ (CB buffer)</td><td>0.172/0.169</td><td>0.151/0.155</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0907" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/7</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>13th</th><th>null</th><th>null</th><th>null</th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>BclAcex+ (PBS buffer) (Bemliese)</td><td>0.201/0.201</td><td>0.201/0.201</td><td></td></tr>
 +
<tr><td>BclActl+ (PBS buffer) (Bemliese)</td><td>0.224/0.223</td><td>0.218/0.215</td><td></td></tr>
 +
<tr><td>BclAcex+ (CB buffer) (Bemliese)</td><td>0.266/0.257</td><td>0.243/0.242</td><td></td></tr>
 +
<tr><td>BclActl+ (CB buffer) (Bemliese)</td><td>0.178/0.177</td><td>0.157/0.149</td><td></td></tr>
 +
<tr><td>BclAcex+ (PBS buffer) (Kimwipe)</td><td>0.203/0.201</td><td>0.200/0.196</td><td></td></tr>
 +
<tr><td>BclActl+ (PBS buffer) (Kimwipe)</td><td>0.221/0.220</td><td>0.224/0.229</td><td></td></tr>
 +
<tr><td>BclAcex+ (CB buffer) (Kimwipe)</td><td>0.273/0.266</td><td>0.244/0.242</td><td></td></tr>
 +
<tr><td>BclActl+ (CB buffer) (Kimwipe)</td><td>0.172/0.172</td><td>0.154/0.154</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0908" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/8</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td>5</td></tr>
 +
<tr><td>2</td><td>9/7 restriction enzyme design BclA scFv X/Pcut</td><td>30</td></tr>
 +
<tr><td>3</td><td>9/7 restriction enzyme design INP CBDcex S/Pcut</td><td>30</td></tr>
 +
<tr><td>4</td><td>9/7 restriction enzyme design INP CBDclos S/Pcut</td><td>30</td></tr>
 +
<tr><td>5</td><td>100bp ladder</td><td>5</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/a/a8/T--Kyoto--9_8_No88_Gel_Extraction_Miyazaki_10_15_12_18_19.1.jpg"><img src="https://static.igem.org/mediawiki/2016/2/2a/T--Kyoto--9_8_No88_Gel_Extraction_Miyazaki_10_15_12_18_19.2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0908" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/8</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>9/5 TransformationINPcex-scFv colony5</td><td>vf 1.25 vr1.25</td><td>10</td><td>KAPA2GFAST/12.5</td><td>25</td></tr>
 +
<tr><td>〃colony6</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃colony7</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃colony8</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>9/5 Transformation INPclos-BclascFv colony1</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃colony2</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃colony5</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃colony6</td><td>=</td><td>=</td><td>=</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/2/26/T--Kyoto--9_8_No89_Colony_PCR_%28typeA%29_Kosaka_10_02_01_52_26.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0908" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/8</h3></span>
 +
<span class="keln_exp"><h4>Western Blotting</h4></span>
 +
<span class="keln_researcher">Ohara, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>lane</th><th>sample</th></tr>
 +
<tr><td>1</td><td>Marker</td></tr>
 +
<tr><td>2</td><td>Sano VLP 0.5&microl</td></tr>
 +
<tr><td>3</td><td>Sano VLP 0.2&microl</td></tr>
 +
<tr><td>4</td><td>Sano VLP 0.1&microl</td></tr>
 +
<tr><td>5</td><td>Sano VLP 0.05&microl</td></tr>
 +
<tr><td>6</td><td>Sano VLP 0.02&microl</td></tr>
 +
<tr><td>7</td><td>Sano VLP 0.05&microl</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/6/64/T--Kyoto--9_8_No90_General_Miyazaki_10_17_05_45_26.jpg">
 +
</div>
 +
<p class="keln_note">1'antibody time/buffer  1:3000 6ml 1h<br>2'antibody time/buffer  1:10000 10ml 1h<br>Blocking 1day<br>transfer 100mA  1h</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0908" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/8</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Insert/&microl</th><th>Vector/&microl</th><th>Ligation High/&microl</th><th>Total</th></tr>
 +
<tr><td>9/8 Gel Extraction BclA scFv X/Pcut /6</td><td>9/8 Gel Extraction INP CBDcex S/Pcut /1.5</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>9/8 Gel Extraction BclA scFv X/Pcut /6</td><td>9/8 Gel Extraction INP CBDcex S/Pcut /1.5</td><td>7.5</td><td>15</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0908" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/8</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamamoto, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>9/8 Ligation Bclascfv-INPCBDcex</td><td>2</td><td>DH5a/20</td><td>LB(CP)</td></tr>
 +
<tr><td>〃BclascFv-INPCBDclos</td><td>2</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>9/7Miniprep BclaCBDcex+8/24 Miniprep J23105</td><td>1</td><td>〃</td><td>LB(CP+AMP)</td></tr>
 +
<tr><td>〃Bcla-CBDclos+8/24 J23105</td><td>1</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>9/7 Miniprep INP-CBDcex+8/24 Miniprep J23105</td><td>1</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>9/7 Miniprep INP-CBDclos+8/24 Miniprep J23105</td><td>1</td><td>〃</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0908" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/8</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>Bcla-CBDcex</td><td>LB(CP)/30ml</td></tr>
 +
<tr><td>Bcla-CBDcontrol</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0908" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/8</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>14th</th><th>null</th><th>null</th><th>null</th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>CBDcex+ (CB buffer)</td><td>0.241/0.236</td><td>0.215/0.212</td><td></td></tr>
 +
<tr><td>CBDctl+ (CB buffer)</td><td>0.182/0.180</td><td>0.159/0.159</td><td></td></tr>
 +
<tr><td>BclAcex+ (CB buffer + Na2CO3)</td><td>0.235/0.235</td><td>0.177/0.177</td><td></td></tr>
 +
<tr><td>BclActl+ (CB buffer+Na2CO3)</td><td>0.227/0.218</td><td>0.155/0.156</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0909" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/9</h3></span>
 +
<span class="keln_exp"><h4>Glycerol Stock</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>sample /ml</th><th>glycerol /ml</th></tr>
 +
<tr><td>BclA control /0.8</td><td>0.2</td></tr>
 +
<tr><td>BclA control /0.8</td><td>0.2</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0909" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/9</h3></span>
 +
<span class="keln_exp"><h4>Glycerol Stock</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>sample /ml</th><th>glycerol /ml</th></tr>
 +
<tr><td>INP control /0.8</td><td>0.2</td></tr>
 +
<tr><td>INP control /0.8</td><td>0.2</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0909" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/9</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0909" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/9</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Iguchi</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>INPcex-scFv colony9</td><td>vf1.25 vr1.25</td><td>8.5</td><td>KAPA2GFAST 12.5</td><td>25</td></tr>
 +
<tr><td>〃colony10</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony11</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>9/5 transformation INPclos-scFv colony1</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃colony5</td><td>〃</td><td>〃</td><td>〃</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/0/02/T--Kyoto--9_9_No96_Colony_PCR_%28typeA%29_Kosaka_10_02_02_29_44.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0909" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/9</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>scFv lc3</td><td>LB()</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0909" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/9</h3></span>
 +
<span class="keln_exp"><h4>Western Blotting</h4></span>
 +
<span class="keln_researcher">Ohara, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>lane</th><th>sample</th></tr>
 +
<tr><td>1</td><td>Marker</td></tr>
 +
<tr><td>2</td><td>Sano VLP 0.5&microl</td></tr>
 +
<tr><td>3</td><td>Sano VLP 1&microl</td></tr>
 +
<tr><td>4</td><td>Sano VLP 0.2&microl</td></tr>
 +
<tr><td>5</td><td>Sano VLP 0.1&microl</td></tr>
 +
<tr><td>6</td><td>Sano VLP 0.05&microl</td></tr>
 +
<tr><td>7</td><td>Sano VLP 0.02&microl</td></tr>
 +
<tr><td>8</td><td>Sano VLP 0.5&microl 95℃ without boiling</td></tr>
 +
<tr><td>9</td><td>only SDS lysis buffer</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/7/7a/T--Kyoto--9_9_No98_General_Miyazaki_10_17_05_52_42.jpg">
 +
</div>
 +
<p class="keln_note">1'antibody time/buffer  1:3000 9ml 1h<br>2'antibody time/buffer  1:10000 10ml 1h<br>Blocking 1h<br>transfer 100mA  1h</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0909" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/9</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>scFv Bcla/1ul</td><td>271.8</td><td>2.09</td><td>2.15</td></tr>
 +
<tr><td>VLP/1ul</td><td>1.67</td><td>1.98</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0909" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/9</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>BclaCBDcexJ23105</td><td>vf1.25 vr1.25</td><td>10</td><td>KAPA2GFAST HSRM withDye/12.5</td><td>25</td></tr>
 +
<tr><td>9/6 Transformation BclaCBDclos J23105</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃INPCBDcex J23105</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃INPCBDcexJ23105</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/1/1b/T--Kyoto--9_9_No100_Colony_PCR_%28typeA%29_Kosaka_10_02_02_51_54.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0909" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/9</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher"></span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>2/24 miniprep Bcla-his-cex colony2</td><td>0.1</td><td>DH5a/20ul</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0909" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/9</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Iguchi</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>9/5 transformation INPcex-BclascFv colony9</td><td>LB(CP)/5ml</td></tr>
 +
<tr><td>〃INPclos-BclascFv colony1</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0909" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/9</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>9/5 transformation Bcla CBDcex J23105 </td><td>vf1.0 vr1.0</td><td>23</td><td>Quick Taq/25</td><td>50</td></tr>
 +
<tr><td>〃BclaCBDclos J23105</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃INPCBDcex J23105</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃INPCBDclos J23105</td><td>〃</td><td>〃</td><td>〃</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0912" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/12</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>9/9miniprep scFv Bcla lc3 </td><td>29ul</td><td></td><td>1</td><td>0</td><td>1</td><td>10×H/5</td><td>5</td><td>9</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0912" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/12</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder/5 </td><td>-</td></tr>
 +
<tr><td>2</td><td>Bcla CBDcex j23105/15①</td><td>789 1140</td></tr>
 +
<tr><td>3</td><td>〃②</td><td>〃</td></tr>
 +
<tr><td>4</td><td>Bcla CBDclos J23105①</td><td>757 1140</td></tr>
 +
<tr><td>5</td><td>〃②</td><td>〃</td></tr>
 +
<tr><td>6</td><td>INP CBDcex J23105①</td><td>1664 1140</td></tr>
 +
<tr><td>7</td><td>〃②</td><td>〃</td></tr>
 +
<tr><td>8</td><td>INP CBDclos J23105①</td><td>1796 1140</td></tr>
 +
<tr><td>9</td><td>〃②</td><td>〃</td></tr>
 +
<tr><td>10</td><td>100bp ladder</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/4/4f/T--Kyoto--9_12_No105_Electrophoresis_Kosaka_10_02_03_10_40.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0912" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/12</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>Bcla CBDcex J23105①</td><td>vf0.5 vr0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>〃②</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃③</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃④</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>Bcla CBDclos J23105①</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃②</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃③</td><td>〃</td><td>〃</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃④</td><td>〃</td><td>〃</td><td>〃</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0912" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/12</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>INP CBDcex J23105</td><td>vf0.5 vr0.5</td><td>11.5</td><td>Quick Taq12.5</td><td>25</td></tr>
 +
<tr><td>INP〃②</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃③</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃④</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃clos①</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃②</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃③</td><td>=</td><td>=</td><td>=</td><td>=</td></tr>
 +
<tr><td>〃④</td><td>=</td><td>=</td><td>=</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0913" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/13</h3></span>
 +
<span class="keln_exp"><h4>Western Blotting</h4></span>
 +
<span class="keln_researcher">Ohara, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>lane</th><th>sample</th></tr>
 +
<tr><td>1</td><td>Marker</td></tr>
 +
<tr><td>2</td><td>Sano VLP 1&microl</td></tr>
 +
<tr><td>3</td><td>Sano VLP 1&microl</td></tr>
 +
<tr><td>4</td><td>Sano VLP 0.2&microl</td></tr>
 +
<tr><td>5</td><td>Sano VLP 0.1&microl</td></tr>
 +
<tr><td>6</td><td>Sano VLP 0.05&microl</td></tr>
 +
<tr><td>7</td><td>Sano VLP 0.02&microl</td></tr>
 +
<tr><td>8</td><td>Sano VLP 0.5&microl 95℃ without boiling</td></tr>
 +
<tr><td>9</td><td>only SDS lysis buffer</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/c/c8/T--Kyoto--9_13_No112_General_Miyazaki_10_17_05_58_13.1.jpg"><img src="https://static.igem.org/mediawiki/2016/3/3c/T--Kyoto--9_13_No112_General_Miyazaki_10_17_05_58_13.2.jpg">
 +
</div>
 +
<p class="keln_note">1'antibody time/buffer  1:3000 9ml 1h<br>2'antibody time/buffer  1:10000 10ml 1h<br>Blocking 1h<br>transfer 100mA  1h<br>30ml TBS-T/blocking reagent 1.5g</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0914" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/14</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>9/9 transformation BclA-CBDcex</td><td>LB(CP)</td></tr>
 +
<tr><td>8/24 transformation BclA-CBDclos</td><td>LB(CP)</td></tr>
 +
<tr><td>8/24 transformation INP-CBDcex</td><td>LB(CP)</td></tr>
 +
<tr><td>8/20 transformation </td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0914" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/14</h3></span>
 +
<span class="keln_exp"><h4>Western Blotting</h4></span>
 +
<span class="keln_researcher">Ohara, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>lane</th><th>sample</th></tr>
 +
<tr><td>1</td><td>Marker</td></tr>
 +
<tr><td>2</td><td>Sano VLP 0.5&microl＋INP-scFv 100&microl </td></tr>
 +
<tr><td>3</td><td>Sano VLP 0.5&microl＋BAN-scFv 100&microl</td></tr>
 +
<tr><td>4</td><td>Sano VLP 0.2&microl ＋BAN-scFv 100&microl</td></tr>
 +
<tr><td>5</td><td>Sano VLP 0.2&microl ＋BAN-scFv 70&microl ＋BANcontrol30&microl </td></tr>
 +
<tr><td>6</td><td>Sano VLP 0.2&microl ＋BAN-scFv 20&microl ＋BANcontrol80&microl </td></tr>
 +
<tr><td>7</td><td>Sano VLP 0.2&microl ＋BANcontrol100&microl</td></tr>
 +
<tr><td>8</td><td>Sano VLP 0.2&microl 95℃ without boiling</td></tr>
 +
<tr><td>9</td><td>Sano VLP 0.5&microl 95℃ without boiling</td></tr>
 +
<tr><td>10</td><td>Marker</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/6/6a/T--Kyoto--9_14_No114_General_Miyazaki_10_17_06_06_07.jpg">
 +
</div>
 +
<p class="keln_note">1'antibody time/buffer  1:1000  10ml / Blocking buffer　overnight<br>2'antibody time/buffer  1:2500 10ml / Blocking buffer 1h<br>Blocking 1h<br>transfer 100mA  1h<br>25ml TBS-T/blocking reagent 1.3g</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0915" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/15</h3></span>
 +
<span class="keln_exp"><h4>Western Blotting</h4></span>
 +
<span class="keln_researcher">Ohara, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>lane</th><th>sample</th></tr>
 +
<tr><td>1</td><td>Marker</td></tr>
 +
<tr><td>2</td><td>Sano VLP 0.5&microl＋INP-scFv 100&microl </td></tr>
 +
<tr><td>3</td><td>Sano VLP 0.5&microl＋BAN-scFv 100&microl</td></tr>
 +
<tr><td>4</td><td>Sano VLP 0.2&microl ＋BAN-scFv 100&microl</td></tr>
 +
<tr><td>5</td><td>Sano VLP 0.2&microl ＋BAN-scFv 70&microl ＋BANcontrol30&microl </td></tr>
 +
<tr><td>6</td><td>Sano VLP 0.2&microl ＋BAN-scFv 20&microl ＋BANcontrol80&microl </td></tr>
 +
<tr><td>7</td><td>Sano VLP 0.2&microl ＋BANcontrol100&microl</td></tr>
 +
<tr><td>8</td><td>Sano VLP 0.2&microl 95℃ without boiling</td></tr>
 +
<tr><td>9</td><td>Sano VLP 0.5&microl 95℃ without boiling</td></tr>
 +
<tr><td>10</td><td>Marker</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/8/8f/T--Kyoto--9_15_No116_General_Miyazaki_10_17_06_11_34.jpg">
 +
</div>
 +
<p class="keln_note">1'antibody time/buffer  1:1000  10ml / Blocking buffer　overnight<br>2'antibody time/buffer  1:2500 10ml / Blocking buffer 1h<br>Blocking 1h<br>transfer 100mA  1h<br>25ml TBS-T/blocking reagent 1.3g</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0915" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/15</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>9/7 BclA scFv 3C5 9/7 2 </td><td>LB(CP) /20mL</td></tr>
 +
<tr><td>9/14 BclA cex (9/9 tranceformation)</td><td>LB(CP) /20mL</td></tr>
 +
<tr><td>9/7 INP scFv 3C5</td><td>LB(CP) /20mL</td></tr>
 +
<tr><td>9/9 BclA scFv 1c3</td><td>LB(CP) /20mL</td></tr>
 +
<tr><td>9/7 INP CTL</td><td>LB(CP) /20mL</td></tr>
 +
<tr><td>9/7 BclA CTL 2</td><td>LB(CP) /20mL</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0915" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/15</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Ohara, Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>BclA CBDclos glycerol stock</td><td>LB(CP)</td></tr>
 +
<tr><td>INP-CBDcex glycerol stock</td><td>LB(CP)</td></tr>
 +
<tr><td>INP-CBDclos glycerol stock</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0916" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/16</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Uchino, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>8/20 J23105 RFP 1/100 </td><td>20</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0919" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/19</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Sassa, Takimoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>2/24 BAN-cex</td><td>1</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>8/5 INP-cex</td><td>1</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>2/8 INP HIsR</td><td>1</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>2/19 BAN-clos</td><td>1</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>8/5 INP-clos</td><td>1</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>2/4 E1010</td><td>1</td><td>DH5α /20</td><td></td></tr>
 +
<tr><td>8/18 BclA-scFv</td><td>1</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0919" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/19</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Sassa, Takimoto</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>9/15 Liquid culture BclA scFv 3C5②</td><td>LB(CP) /20</td></tr>
 +
<tr><td>9/19 Liquid culture　Bcla CTL</td><td>LB(CP) /20</td></tr>
 +
<tr><td>9/15 Liquid culture INP-scFv 3C5</td><td>LB(CP) /20</td></tr>
 +
<tr><td>9/15 Liquid culture INP CTL</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0920" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/20</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>9/19 liquid culture BclA-scFv 3C5-2</td><td>LB(CP)</td></tr>
 +
<tr><td>9/19 liquid culture BclA-ctl</td><td>LB(CP)</td></tr>
 +
<tr><td>9/19 liquid culture INP-scFv 3C5</td><td>LB(CP)</td></tr>
 +
<tr><td>9/19 liquid culture INP-ctl</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0920" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/20</h3></span>
 +
<span class="keln_exp"><h4>Western Blotting</h4></span>
 +
<span class="keln_researcher">Ohara, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>lane</th><th>sample</th></tr>
 +
<tr><td>Marker</td><td>M.Marker</td></tr>
 +
<tr><td>1</td><td>INPNC-6His-scFv 300 &microl＋VLP3 &microl</td></tr>
 +
<tr><td>2</td><td>INPNC-6His-scFv 300 &microl＋PBS3 &microl </td></tr>
 +
<tr><td>3</td><td>INPNC-6His-ctl300 &microl＋VLP3 &microl </td></tr>
 +
<tr><td>4</td><td>BclA-6His-scFv 300 &microl＋VLP3 &microl </td></tr>
 +
<tr><td>5</td><td>BclA-6His-scFv 300 &microl＋PBS3 &microl </td></tr>
 +
<tr><td>6</td><td>BclA-6His-ctl 300 &microl＋VLP3 &microl </td></tr>
 +
<tr><td>7</td><td>VLP0.5 &microl</td></tr>
 +
<tr><td>8</td><td>VLP0.2 &microl</td></tr>
 +
<tr><td>9</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/4/48/T--Kyoto--9_20_No124_General_Miyazaki_10_17_06_17_54.jpg">
 +
</div>
 +
<p class="keln_note">Transfer ;60 min 100 mA(CC)<br>Blocking ;1h<br>1’antibody ;anti-NoV pAb(mouse)<br>1:1000 with blocking buffer  O/N<br>2’antibody ;anti-mouse HRP　<br>1:2500 with blocking buffer 1 h<br>Detection time　80 sec<br>(ECL Prime)</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0920" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/20</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>BclA-clos glycerol stock</td><td>-</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>INP-clos glycerol stock</td><td>-</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0921" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/21</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Predenature</td><td>95℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>98℃</td><td>20sec</td><td>30</td></tr>
 +
<tr><td>Annealing</td><td>65℃</td><td>15sec</td><td>30</td></tr>
 +
<tr><td>Extension</td><td>72℃</td><td>30sec</td><td>30</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>3min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>8℃</td><td>-</td><td>-</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0921" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/21</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/8 miniprep INP-His9 /0.2</td><td>VF /0.75</td><td>m.scFv Rv /0.75</td><td>-</td><td>10.8</td><td>KAPA HiFi Ready Mix /12.5</td><td>25</td></tr>
 +
<tr><td>2/8 miniprep INP-His9 /0.2</td><td>m.scFv Fw /0.75</td><td>VR /0.75</td><td>-</td><td>10.8</td><td>KAPA HiFi Ready Mix /12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">VF 10&microM      VR 10&microM</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0921" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/21</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td>5</td></tr>
 +
<tr><td>2</td><td>100bp ladder </td><td>5</td></tr>
 +
<tr><td>3</td><td>9/21 PCR INP-His9-1</td><td>25</td></tr>
 +
<tr><td>4</td><td>9/21 PCR INP-His9-2</td><td>25</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/5/53/T--Kyoto--9_21_No129_Gel_Extraction_Notsu_10_03_01_11_36.1.jpg"><img src="https://static.igem.org/mediawiki/2016/b/b2/T--Kyoto--9_21_No129_Gel_Extraction_Notsu_10_03_01_11_36.2.jpg">
 +
</div>
 +
<img src="https://static.igem.org/mediawiki/2016/b/b2/T--Kyoto--9_21_No129_Gel_Extraction_Notsu_10_03_01_11_36.2.jpg">
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0921" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/21</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Ohara, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>9/19 Liquid culture BAN-scFv 3C5②</td><td>LB(CP)</td></tr>
 +
<tr><td>9/19 Liquid culture BAN-CTL </td><td>LB(CP)</td></tr>
 +
<tr><td>9/19 Liquid culture INP-scFv 3C5</td><td>LB(CP)</td></tr>
 +
<tr><td>9/19 Liquid culture INP-CTL</td><td>LB(CP)</td></tr>
 +
<tr><td>9/15 BAN- scFv 1C3</td><td>LB(CP)</td></tr>
 +
<tr><td>9/15 BAN-CBDcex</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0921" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/21</h3></span>
 +
<span class="keln_exp"><h4>Western Blotting</h4></span>
 +
<span class="keln_researcher">Ohara, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>lane</th><th>sample</th></tr>
 +
<tr><td>Marker</td><td>M.Marker</td></tr>
 +
<tr><td>1</td><td>INPNC-6His-scFv 200 &microl＋VLP2 &microl</td></tr>
 +
<tr><td>2</td><td>INPNC-6His-scFv 200 &microl＋PBS2&microl(NC)</td></tr>
 +
<tr><td>3</td><td>INPNC-6His-ctl200 &microl＋VLP2 &microl(NC)</td></tr>
 +
<tr><td>4</td><td>BclA-6His-scFv200 ul(Vector pSB3C5)＋VLP2 ul</td></tr>
 +
<tr><td>5</td><td>BclA-6His-scFv 200 &microl＋VLP2 &microl (NC)</td></tr>
 +
<tr><td>6</td><td> BclA-6His-scFv(Vector pSB1C3)＋VLP2 ul</td></tr>
 +
<tr><td>7</td><td> BclA-6His-CBDcex 200 ul ＋ VLP2 ul (NC)</td></tr>
 +
<tr><td>8</td><td>PBS200 ul ＋VLP2 ul (NC)</td></tr>
 +
<tr><td>9</td><td>VLP1 ul (PC) </td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/1/16/T--Kyoto--9_21_No131_General_Miyazaki_10_17_06_47_39.jpg">
 +
</div>
 +
<p class="keln_note">Transfer ;60 min 100 mA(CC)<br>Blocking ;1h<br>1’antibody ;anti-NoV pAb(mouse)<br>1:1000 with blocking buffer  O/N<br>2’antibody ;anti-mouse HRP　<br>1:2500 with blocking buffer 1 h<br>Detection time　60 sec<br>(ECL Prime)</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0921" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/21</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Sample/(&microl)</th><th>Concentration(&microg/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>9/21 PCR INP-His9-1</td><td>19.3</td><td>1.63</td><td>0.54</td></tr>
 +
<tr><td>9/21 PCR INP-His9-2</td><td>48.6</td><td>1.82</td><td>0.24</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0921" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/21</h3></span>
 +
<span class="keln_exp"><h4>PCR (Steps)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Steps</th><th>Temparature</th><th>Time</th><th>Cycle</th></tr>
 +
<tr><td>Predenature</td><td>95℃</td><td>5min</td><td>1</td></tr>
 +
<tr><td>Denature</td><td>98℃</td><td>20sec</td><td>30</td></tr>
 +
<tr><td>Annealing</td><td>65℃</td><td>15sec</td><td>30</td></tr>
 +
<tr><td>Extension</td><td>72℃</td><td>30sec</td><td>30</td></tr>
 +
<tr><td>Final Elongation</td><td>72℃</td><td>3min</td><td>1</td></tr>
 +
<tr><td>Final Holding</td><td>8℃</td><td>-</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0921" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/21</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>9/21 Gel Extract INP-His9-1 /1&microl+9/21 Gel Extract INP-His9-2 /0.5&microl</td><td>VF /0.75  </td><td>VR /0.75</td><td>-</td><td>9.5</td><td>KAPA HiFi /12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0922" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/22</h3></span>
 +
<span class="keln_exp"><h4>growth carve</h4></span>
 +
<span class="keln_researcher">Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>sample</th><th>0h</th><th>0.5h</th><th>1h</th><th>1.5h</th><th>2h</th><th>3h</th><th>4h</th><th>5h</th><th>6h</th><th>7h</th><th>8h</th></tr>
 +
<tr><td>BAM scFv　①</td><td>-0.048</td><td>-0.047</td><td>-0.048</td><td>1.245</td><td>-0.317</td><td>-0.308</td><td>-0.149</td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>BAM scFv　②</td><td>-0.036</td><td>1.512</td><td>-0.830</td><td>-0.309</td><td>-0.314</td><td>-0.127</td><td></td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>INP scFv　①</td><td>-0.047</td><td>-0.050</td><td>-0.644</td><td>1.045</td><td>-0.307</td><td>-0.153</td><td>-0.142</td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>INP scFv　②</td><td>-0.038</td><td>-0.o25</td><td>-0.186</td><td>-0.280</td><td>0.000</td><td>-0.289</td><td></td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>BAN CBDcex　①</td><td>-0.045</td><td>-0.048</td><td>-0.044</td><td>1.089</td><td>-0.322</td><td>-0.284</td><td>-0.296</td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>BAN CBDcex　②</td><td>-0.034</td><td>1.204</td><td>-1.057</td><td>-0.311</td><td>0.268</td><td>-0.285</td><td></td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>BAN CBDclos　①</td><td>0.035</td><td>-0.037</td><td>-0.035</td><td>1.261</td><td>-0.323</td><td>-0.160</td><td>-0.287</td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>BAN CBDclos　②</td><td>-0.018</td><td>1.306</td><td>-0.050</td><td>-0.282</td><td>-0.128</td><td>-0.293</td><td></td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>INP CBDcex　①</td><td>-0.020</td><td>-0.027</td><td>-0.016</td><td>1.245</td><td>-0.295</td><td>-0.263</td><td>-0.288</td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>INP CBDcex　②</td><td>-0.027</td><td>1.608</td><td>0.016</td><td>-0.287</td><td>-0.224</td><td>-0.294</td><td></td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>INP CBDclos　①</td><td>-0.034</td><td>-0.034</td><td>-0.032</td><td>1.026</td><td>-0.318</td><td>-0.235</td><td>-0.270</td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>INP CBDclos　②</td><td>-0.026</td><td>1.415</td><td>0.012</td><td>-0.297</td><td>-0.047</td><td>-0.272</td><td></td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>BAN CTL 　①</td><td>-0.039</td><td>-0.039</td><td>-0.036</td><td>1.274</td><td>-0.314</td><td>-0.230</td><td>-0.271</td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>BAN CTL 　②</td><td>-0.056</td><td>1.530</td><td>-0.060</td><td>-0.284</td><td>-0.084</td><td>-3.11</td><td></td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>INP CTL　①</td><td>-0.042</td><td>-0.043</td><td>-0.042</td><td>1.134</td><td>-0.235</td><td>-0.181</td><td>-0.296</td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>INP CTL　②</td><td>-0.042</td><td>-1.488</td><td>-0.030</td><td>-0.288</td><td>-0.109</td><td>-0.295</td><td></td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>E1010　①</td><td>-0.041</td><td>-0.040</td><td>-0.036</td><td>1.171</td><td>-0.322</td><td>-0.252</td><td></td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>E1010　②</td><td>-0.030</td><td>1.563</td><td>-0.061</td><td>-0.267</td><td>0.286</td><td>-0.267</td><td></td><td></td><td></td><td></td><td></td></tr>
 +
<tr><td>blank ①/②</td><td></td><td></td><td></td><td> /-0.307</td><td>-0.306/-0.305</td><td>-0.307/-0.271</td><td>0.306/</td><td></td><td></td><td></td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Notsu -->
 +
<div class="keln_container">
 +
<a name="0922" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/22</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Sassa, Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>9/21 liquid culture BAN-CBDcex</td><td>LB(AMP)</td></tr>
 +
<tr><td>9/15 liquid culture INP-ctl</td><td>LB(AMP)</td></tr>
 +
<tr><td>9/15 liquid culture INP-CBDcex</td><td>LB(AMP)</td></tr>
 +
<tr><td>9/15 liquid culture INP-CBDclos</td><td>LB(AMP)</td></tr>
 +
<tr><td>9/15 liquid culture BAN-scFv 3C5</td><td>LB(AMP)</td></tr>
 +
<tr><td>9/15 liquid culture INP-scFv 3C5</td><td>LB(AMP)</td></tr>
 +
<tr><td>9/19 liquid culture BAN-CBDclos</td><td>LB(AMP)</td></tr>
 +
<tr><td>9/15 liquid culture BAN-ctl</td><td>LB(AMP)</td></tr>
 +
<tr><td>9/16 transformation J23105-RFP </td><td>LB(AMP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0923" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/23</h3></span>
 +
<span class="keln_exp"><h4>SEM</h4></span>
 +
<span class="keln_researcher">Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>number</th><th>sample(&microl)</th><th>VLP(&microl)</th><th>PBS</th><th>centrifugal force</th></tr>
 +
<tr><td>1</td><td>INP scFv /25</td><td>2.5</td><td>0</td><td>×3000g</td></tr>
 +
<tr><td>2</td><td>INP CTL /25</td><td>2.5</td><td>0</td><td>×3000g</td></tr>
 +
<tr><td>3</td><td>BclA scFv /25</td><td>2.5</td><td>0</td><td>×3000g</td></tr>
 +
<tr><td>4</td><td>BclA CTL /25</td><td>2.5</td><td>0</td><td>×3000g</td></tr>
 +
<tr><td>5</td><td>BclA scFv /25</td><td>0</td><td>2.5</td><td>×3000g</td></tr>
 +
<tr><td>6</td><td>INP scFv /25</td><td>2.5</td><td>0</td><td>×200g</td></tr>
 +
<tr><td>7</td><td>INP CTL /25</td><td>2.5</td><td>0</td><td>×200g</td></tr>
 +
<tr><td>8</td><td>BclA scFv /25</td><td>2.5</td><td>0</td><td>×200g</td></tr>
 +
<tr><td>9</td><td>BclA CTL /25</td><td>2.5</td><td>0</td><td>×200g</td></tr>
 +
<tr><td>10</td><td>BclA scFv /25</td><td>0</td><td>2.5</td><td>×200g</td></tr>
 +
<tr><td>11</td><td>0</td><td>2.5</td><td>0</td><td>×200g</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0923" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/23</h3></span>
 +
<span class="keln_exp"><h4>RNA extraction</h4></span>
 +
<span class="keln_researcher">Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>number</th><th>sample name</th></tr>
 +
<tr><td>1</td><td>BclA-CBD cex</td></tr>
 +
<tr><td>2</td><td>BclA-CBDclos</td></tr>
 +
<tr><td>3</td><td>INP-CBDcex</td></tr>
 +
<tr><td>4</td><td>INP-CBDclos</td></tr>
 +
<tr><td>5</td><td>BclA-scFv</td></tr>
 +
<tr><td>6</td><td>INP-scFv</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0923" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/23</h3></span>
 +
<span class="keln_exp"><h4>growth carve</h4></span>
 +
<span class="keln_researcher"></span>
 +
<table class="keln_table">
 +
<tr><th>sample</th><th>0h</th><th>0.5h</th><th>1h</th><th>1.5h</th><th>2h</th><th>3h</th><th>4h</th><th>5h</th><th>6h</th><th>7h</th><th>8h</th><th>9h</th><th>10h</th></tr>
 +
<tr><td>9/19 Liquid culture BAN scFv①</td><td>0.003</td><td>0.001</td><td>0.000</td><td>0.008</td><td>0.015</td><td>0.009</td><td>0.013</td><td>0.016</td><td>0.015</td><td>0.022</td><td>0.062</td><td>0.020</td><td>0.027</td></tr>
 +
<tr><td>9/19 Liquid culture BAN scFv②</td><td>0.013</td><td>0.012</td><td>0.000</td><td>0.000</td><td>0.000</td><td>0.000</td><td>0.001</td><td>0.011</td><td>0.011</td><td>0.015</td><td>0.022</td><td>0.027</td><td>0.057</td></tr>
 +
<tr><td>9/19 Liquid culture INP scFv①</td><td>0.012</td><td>0.000</td><td>0.000</td><td>0.011</td><td>0.006</td><td>0.006</td><td>0.007</td><td>0.008</td><td>0.013</td><td>0.025</td><td>0.036</td><td>0.098</td><td>0.076</td></tr>
 +
<tr><td>9/19 Liquid culture INP scFv②</td><td>0.007</td><td>0.005</td><td>0.000</td><td>0.002</td><td>0.003</td><td>0.002</td><td>0.012</td><td>0.012</td><td>0.019</td><td>0.029</td><td>0.034</td><td>0.047</td><td>0.072</td></tr>
 +
<tr><td>9/19 Liquid culture BAN cex①</td><td>0.014</td><td>0.005</td><td>0.010</td><td>0.008</td><td>0.011</td><td>0.009</td><td>0.014</td><td>0.014</td><td>0.016</td><td>0.029</td><td>0.038</td><td>0.043</td><td>0.083</td></tr>
 +
<tr><td>9/19 Liquid culture BAN cex②</td><td>0.011</td><td>0.011</td><td>0.010</td><td>0.008</td><td>0.004</td><td>0.004</td><td>0.020</td><td>0.021</td><td>0.036</td><td>0.047</td><td>0.065</td><td>0.087</td><td>0.109</td></tr>
 +
<tr><td>9/19 Liquid culture BAN clos①</td><td>0.000</td><td>0.000</td><td>0.000</td><td>0.002</td><td>0.009</td><td>0.003</td><td>0.009</td><td>0.014</td><td>0.019</td><td>0.014</td><td>0.016</td><td>0.024</td><td>0.046</td></tr>
 +
<tr><td>9/19 Liquid culture BAN clos②</td><td>0.005</td><td>0.007</td><td>0.000</td><td>0.000</td><td>0.000</td><td>0.003</td><td>0.012</td><td>0.022</td><td>0.020</td><td>0.035</td><td>0.055</td><td>0.086</td><td>0.117</td></tr>
 +
<tr><td>9/19 Liquid culture INP cex①</td><td>0.187</td><td>0.000</td><td>0.001</td><td>0.007</td><td>0.011</td><td>0.006</td><td>0.013</td><td>0.016</td><td>0.020</td><td>0.023</td><td>0.037</td><td>0.048</td><td>0.068</td></tr>
 +
<tr><td>9/19 Liquid culture INP cex②</td><td>0.002</td><td>0.009</td><td>0.000</td><td>0.001</td><td>0.001</td><td>0.002</td><td>0.015</td><td>0.017</td><td>0.023</td><td>0.036</td><td>0.053</td><td>0.080</td><td>0.111</td></tr>
 +
<tr><td>9/19 Liquid culture INP clos①</td><td>0.006</td><td>0.000</td><td>0.000</td><td>0.023</td><td>0.011</td><td>0.003</td><td>0.011</td><td>0.013</td><td>0.016</td><td>0.022</td><td>0.033</td><td>0.058</td><td>0.068</td></tr>
 +
<tr><td>9/19 Liquid culture INP clos②</td><td>0.029</td><td>0.028</td><td>0.000</td><td>0.009</td><td>0.000</td><td>0.016</td><td>0.001</td><td>0.007</td><td>0.008</td><td>0.014</td><td>0.020</td><td>0.029</td><td>0.074</td></tr>
 +
<tr><td>9/19 Liquid culture BAN CTL①</td><td>0.004</td><td>0.000</td><td>0.000</td><td>0.000</td><td>0.002</td><td>0.000</td><td>0.002</td><td>0.012</td><td>0.015</td><td>0.012</td><td>0.027</td><td>0.055</td><td>0.081</td></tr>
 +
<tr><td>9/19 Liquid culture BAN CTL②</td><td>0.002</td><td>0.002</td><td>0.001</td><td>0.000</td><td>0.001</td><td>0.005</td><td>0.017</td><td>0.024</td><td>0.031</td><td>0.042</td><td>0.059</td><td>0.073</td><td>0.113</td></tr>
 +
<tr><td>9/19 Liquid culture INP CTL①</td><td>0.011</td><td>0.000</td><td>0.032</td><td>0.013</td><td>0.017</td><td>0.010</td><td>0.015</td><td>0.033</td><td>0.031</td><td>0.035</td><td>0.037</td><td>0.019</td><td>0.026</td></tr>
 +
<tr><td>9/19 Liquid culture INP CTL②</td><td>0.013</td><td>0.020</td><td>0.000</td><td>0.001</td><td>0.000</td><td>0.003</td><td>0.009</td><td>0.016</td><td>0.021</td><td>0.026</td><td>0.044</td><td>0.059</td><td>0.136</td></tr>
 +
<tr><td>9/19 Liquid culture E1010①</td><td>0.015</td><td>0.016</td><td>0.014</td><td>0.011</td><td>0.016</td><td>0.018</td><td>0.032</td><td>0.066</td><td>0.108</td><td>0.173</td><td>0.258</td><td>0.361</td><td>0.393</td></tr>
 +
<tr><td>9/19 Liquid culture E1010②</td><td>0.064</td><td>0.010</td><td>0.007</td><td>0.011</td><td>0.012</td><td>0.019</td><td>0.029</td><td>0.041</td><td>0.061</td><td>0.097</td><td>0.150</td><td>0.180</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0923" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/23</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder/5</td><td></td></tr>
 +
<tr><td>2</td><td>9/21 OEPCR/25</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/6/6f/T--Kyoto--9_23_No139_Electrophoresis_Miyazaki_10_06_03_28_24.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0923" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/23</h3></span>
 +
<span class="keln_exp"><h4>OEPCR</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>sample1 /&microl</th><th>sample2 /&microl</th><th>primer Fw /&microl</th><th>primer Rv /&microl</th><th>milliQ /&microl</th><th>KAPA HiFi /&microl</th><th>total</th></tr>
 +
<tr><td>9/21 Gel Extract1 /1</td><td>9/21 Gel Extract2 /0.5</td><td>VF/ 0.75</td><td>VR /0.75</td><td>9.5</td><td>12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0923" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/23</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder/3</td><td></td></tr>
 +
<tr><td>2</td><td>9/23 OEPCR1 /25</td><td></td></tr>
 +
<tr><td>3</td><td>9/23 OEPCR2 /25</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/8/82/T--Kyoto--9_23_No141_Electrophoresis_Miyazaki_10_06_03_34_18.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0923" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/23</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/8 miniprep INP-His-scFv9 /0.4</td><td>m.scFv Fw/1.25</td><td>VR/1.25</td><td>KAPA2GFAST/12.5</td><td>9.6</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/8 miniprep INP-His-scFv9 /0.4</td><td>m.scFv Fw/1.25</td><td>VR/1.25</td><td>KAPA2GFAST/12.5</td><td>9.6</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/8 miniprep INP-His-scFv9 /0.4</td><td>VF/1.25</td><td>m.scFv Rv/1.25</td><td>KAPA2GFAST/12.5</td><td>9.6</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/8 miniprep INP-His-scFv9 /0.4</td><td>VF/1.25</td><td>m.scFv Rv/1.25</td><td>KAPA2GFAST/12.5</td><td>9.6</td><td>0</td><td>25</td></tr>
 +
<tr><td>9/23 Gel Extraction INP-scFv m.PCR/1.5</td><td>VF/1.25</td><td>VR/1.25</td><td>KAPA2GFAST/12.5</td><td>8.5</td><td>0</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0926" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/26</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder/4</td><td></td></tr>
 +
<tr><td>2</td><td>9/23PCR1 /25</td><td></td></tr>
 +
<tr><td>3</td><td>9/23PCR2 /25</td><td></td></tr>
 +
<tr><td>4</td><td>9/23PCR3 /25</td><td></td></tr>
 +
<tr><td>5</td><td>9/23PCR4 /25</td><td></td></tr>
 +
<tr><td>6</td><td>9/23PCR5 /25</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/6/68/T--Kyoto--9_26_No143_Electrophoresis_Miyazaki_10_06_03_46_00.1.jpg"><img src="https://static.igem.org/mediawiki/2016/1/18/T--Kyoto--9_26_No143_Electrophoresis_Miyazaki_10_06_03_46_00.2.jpg">
 +
</div>
  
−
<!-- Table Generated by KELN Author: Sukegawa -->
+
<!------------ Table END ------------>
−
 
+
<!-- Table Generated by KELN Author: Miyazaki -->
 
<div class="keln_container">
 
<div class="keln_container">
 +
<a name="0926" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/26</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>9/26 electrophoresis lane 4,5</td><td></td><td>0.357g</td></tr>
 +
<tr><td>9/26 electrophoresis lane 2,3</td><td></td><td>0.354g</td></tr>
 +
</table>
 +
</div>
  
−
<a name="0101" class = "kyoto-jump"></a>
+
<!------------ Table END ------------>
−
 
+
<!-- Table Generated by KELN Author: Miyazaki -->
−
<span class="keln_date"><h3>1/1</h3></span>
+
<div class="keln_container">
−
 
+
<a name="0926" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/26</h3></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>9/26 Gel Extract lane4,5 /2</td><td>VF/0.75</td><td>VR/0.75</td><td>2×KAPA HiFi Hotstart ReadyMix/12.5</td><td>7</td><td></td><td>25</td></tr>
 +
<tr><td>9/26 Gel Extract lane2,3 /2</td><td>VF/0.75</td><td>VR/0.75</td><td>2×KAPA HiFi Hotstart ReadyMix/12.5</td><td>7</td><td></td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0926" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/26</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>INP-scFv/30ml</td><td>LB</td></tr>
 +
<tr><td>INP-CTL/30ml</td><td>LB</td></tr>
 +
<tr><td>BAN-scFv/30ml</td><td>LB</td></tr>
 +
<tr><td>Ban CTL/30ml</td><td>LB</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0926" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/26</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>Bcla-cex 8/5 miniprep</td><td></td><td>BL21</td><td>LB(CP)</td></tr>
 +
<tr><td>Bcla-CTL 8/4 miniprep</td><td></td><td>BL21</td><td>LB(CP)</td></tr>
 +
<tr><td>Bcla-cex 8/5 miniprep</td><td></td><td>BL21</td><td>LB(CP)</td></tr>
 +
<tr><td>Bcla-CTL 8/4 miniprep</td><td></td><td>BL21</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0926" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/26</h3></span>
 +
<span class="keln_exp"><h4>RNA extraction</h4></span>
 +
<span class="keln_researcher">Isoda</span>
 +
<table class="keln_table">
 +
<tr><th>number</th><th>sample</th><th>conc.&microg/&microl</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>4</td><td>BclAcex</td><td>74.73</td><td>1.94</td><td>1.61</td></tr>
 +
<tr><td>2</td><td>INPcex</td><td>83</td><td>1.97</td><td>1.38</td></tr>
 +
<tr><td>5</td><td>INPclos</td><td>120.21</td><td>1.99</td><td>1.94</td></tr>
 +
<tr><td>3</td><td>INPclos</td><td>58.23</td><td>1.98</td><td>1.84</td></tr>
 +
<tr><td>6</td><td>BclAscFV</td><td>74.76</td><td>1.96</td><td>1.84</td></tr>
 +
<tr><td>7</td><td>INPscFv</td><td>80.6</td><td>1.97</td><td>1.97</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0927" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/27</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>BclA-cex /250ml</td><td>LB(CP)</td></tr>
 +
<tr><td>BclA-CTL /250ml</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="0927" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/27</h3></span>
 +
<span class="keln_exp"><h4>growth carve</h4></span>
 +
<span class="keln_researcher">Sassa, Takimoto</span>
 +
<table class="keln_table">
 +
<tr><th>sample</th><th>0h</th><th>0.5h</th><th>1h</th><th>1.5h</th><th>2h</th><th>3h</th><th>4h</th><th>5h</th><th>6h</th><th>7h</th><th>8h</th></tr>
 +
<tr><td>9/21Liquid culture BAN scFv</td><td>0.113</td><td>0.170</td><td>0.266</td><td>0.408</td><td>0.599</td><td>0.893</td><td>0.164(1.64)</td><td>0.169(1.69)</td><td>0.232(2.32)</td><td>0.234(2.34)</td><td>0.221(2.21)</td></tr>
 +
<tr><td>9/22Liquid culture BAN cex</td><td>0.109</td><td>0.147</td><td>0.219</td><td>0.374</td><td>0.515</td><td>0.873</td><td>0.146(1.46)</td><td>0.160(1.60)</td><td>0.176(1.76)</td><td>0.231(2.31)</td><td>0.260(2.60)</td></tr>
 +
<tr><td>9/19Liquid culture BAN clos</td><td>0.097</td><td>0.109</td><td>0.184</td><td>0.338</td><td>0.486</td><td>0.744</td><td>0.130(1.30)</td><td>0.173(1.73)</td><td>0.199(1.99)</td><td>0.203(2.03)</td><td>0.274(2.74)</td></tr>
 +
<tr><td>9/21Liquid culture  BAN CTL</td><td>0.106</td><td>0.170</td><td>0.252</td><td>0.304</td><td>0.553</td><td>0.797</td><td>0.177(1.77)</td><td>0.155(1.55)</td><td>0.216(2.16)</td><td>0.236(2.36)</td><td>0.257(2.57)</td></tr>
 +
<tr><td>9/21Liquid culture INP scFv</td><td>0.088</td><td>0.083</td><td>0.101</td><td>0.122</td><td>0.299</td><td>0.409</td><td>0.073(0.73)</td><td>0.121(1.21)</td><td>0.140(1.40)</td><td>0.143(1.43)</td><td>0.195(1.95)</td></tr>
 +
<tr><td>9/15Liquid culture INP cex</td><td>0.102</td><td>0.116</td><td>0.145</td><td>0.207</td><td>0.320</td><td>0.626</td><td>0.170(1.70)</td><td>0.143(1.43)</td><td>0.176(1.76)</td><td>0.188(1.88)</td><td>0.238(2.38)</td></tr>
 +
<tr><td>9/22Liquid culture INP clos</td><td>---</td><td>0.133</td><td>0.209</td><td>0.373</td><td>0.572</td><td>0.930</td><td>0.156(1.56)</td><td>0.201(2.01)</td><td>0.246(2.46)</td><td>0.237(2.37)</td><td>0.293(2.93)</td></tr>
 +
<tr><td>9/21Liquid culture INP CTL</td><td>0.112</td><td>0.155</td><td>0.237</td><td>0.409</td><td>0.582</td><td>0.955</td><td>0.153(1.53)</td><td>0.209(2.09)</td><td>0.268(2.68)</td><td>0.288(2.88)</td><td>0.274(2.74)</td></tr>
 +
<tr><td>9/21Liquid culture E1010</td><td>0.073</td><td>0.091</td><td>0.149</td><td>0.265</td><td>0.396</td><td>0.706</td><td>0.157(1.57)</td><td>0.182(1.82)</td><td>0.222(2.22)</td><td>0.222(2.22)</td><td></td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">rpm 160<br>total10mL<br>CP　2mL/1L<br>4h~ 10倍希釈</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0927" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/27</h3></span>
 +
<span class="keln_exp"><h4>Western Blotting</h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>lane</th><th>sample</th></tr>
 +
<tr><td>Marker</td><td>M.Marker(Wako)</td></tr>
 +
<tr><td>1</td><td>BclA6His-scFv 300 &microl＋supernatant of NoVLP 3 &microl (washed by PBS) </td></tr>
 +
<tr><td>2</td><td>BclA6His ctl 300 &microl＋supernatant of NoVLP 3 &microl (NC) (washed by PBS) </td></tr>
 +
<tr><td>3</td><td>INPNC6His-scFv 300 &microl＋supernatant of NoVLP 3 &microl (washed by PBS) </td></tr>
 +
<tr><td>4</td><td>INPNC6His ctl 300 &microl＋supernatant of NoVLP 3 &microl (NC) (washed by PBS) </td></tr>
 +
<tr><td>5</td><td>BclA6His-scFv 300 &microl＋supernatant of NoVLP 3&microl (washed by PBST (0.1% tween20)) </td></tr>
 +
<tr><td>6</td><td>BclA6His ctl 300 &microl＋supernatant of NoVLP 3&microl (NC) (washed by PBST (0.1% tween20)) </td></tr>
 +
<tr><td>7</td><td>INPNC6His-scFv 300 &microl＋supernatant of NoVLP 3&microl (washed by PBST (0.1% tween20)) </td></tr>
 +
<tr><td>8</td><td>INPNC6His ctl 300 &microl＋supernatant of NoVLP 3&microl (NC) (washed by PBST (0.1% tween20)) </td></tr>
 +
<tr><td>9</td><td>PBS 300&microl＋supernatant of NoVLP 3 &microl (NC) (washed by PBS)</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/3/3d/T--Kyoto--9_27_No149_General_Miyazaki_10_17_06_53_43.png">
 +
</div>
 +
<p class="keln_note">Transfer ;60 min 100 mA(CC)<br>Blocking ;1h<br>1’antibody ;anti-NoV pAb(mouse)<br>1:1000 with blocking buffer  O/N<br>2’antibody ;anti-mouse HRP　<br>1:2500 with blocking buffer 1 h<br>Detection time　60 sec<br>(ECL Prime)</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0927" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/27</h3></span>
 +
<span class="keln_exp"><h4>Western Blotting</h4></span>
 +
<span class="keln_researcher">Ohara, Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>lane</th><th>sample</th></tr>
 +
<tr><td>Marker</td><td>M.Marker(Wako) 5&microl</td></tr>
 +
<tr><td>1</td><td>BBa_E1010(NC) </td></tr>
 +
<tr><td>2</td><td>BclA-6His-CBDclos (17 kDa) </td></tr>
 +
<tr><td>3</td><td>INPNC-6His-CBDclos(47 kDa) </td></tr>
 +
<tr><td>4</td><td>BclA-6His-CBDcex  (17 kDa) </td></tr>
 +
<tr><td>5</td><td>INPNC-6His-CBDcex (47 kDa)  </td></tr>
 +
<tr><td>6</td><td>BclA-6His-scFv (33 kDa) </td></tr>
 +
<tr><td>7</td><td>INPNC-6His-scFv (63 kDa) </td></tr>
 +
<tr><td>8</td><td>BBa_K875004 (PC)  Lpp-OmpA-scFv-6His (43kDa) </td></tr>
 +
<tr><td>Marker</td><td> M.Marker(Wako) 3&microl</td></tr>
 +
<tr><td>9</td><td>BBa_K875004 (PC)  Lpp-OmpA-scFv-6His(43 kDa) </td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/2/2a/T--Kyoto--9_27_No149_General_Miyazaki_10_17_07_01_58.png">
 +
</div>
 +
<p class="keln_note">Used membrane fraction from 100ml culture at logarithmic growth phase.<br><br>Transfer ;60min 100mA(CC)<br>Blocking ;1h<br>1’antibody ;anti6His(mouse)<br>1:1000 with blocking buffer  O/N<br>2’antibody ;anti-mouse HRP　<br>1:2500 with TBST 1h<br>Detection time　380sec<br>(ECL Prime)</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0927" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/27</h3></span>
 +
<span class="keln_exp"><h4>Reverse Transcription</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>number</th><th>Template/&microl</th><th>gDNA wipeout buffer/&microl</th><th>RNAase free water/&microl</th><th>total/&microl</th></tr>
 +
<tr><td>A1</td><td>BclAcex/12</td><td>2</td><td>0</td><td>14</td></tr>
 +
<tr><td>A2</td><td>BclAclos/12</td><td>2</td><td>0</td><td>14</td></tr>
 +
<tr><td>A3</td><td>INPcex/8.32</td><td>2</td><td>3.68</td><td>14</td></tr>
 +
<tr><td>A4</td><td>INPclos/12</td><td>2</td><td>0</td><td>14</td></tr>
 +
<tr><td>A5</td><td>BclAscFv/12</td><td>2</td><td>0</td><td>14</td></tr>
 +
<tr><td>A6</td><td>INPscFv/12</td><td>2</td><td>0</td><td>14</td></tr>
 +
<tr><td>B1</td><td>BclAcex/12</td><td>2</td><td>0</td><td>14</td></tr>
 +
<tr><td>B2</td><td>BclAclos/12</td><td>2</td><td>0</td><td>14</td></tr>
 +
<tr><td>B3</td><td>INPcex/8.32</td><td>2</td><td>3.68</td><td>14</td></tr>
 +
<tr><td>B4</td><td>INPclos/12</td><td>2</td><td>0</td><td>14</td></tr>
 +
<tr><td>B5</td><td>BclAscFv/12</td><td>2</td><td>0</td><td>14</td></tr>
 +
<tr><td>B6</td><td>INPscFv/12</td><td>2</td><td>0</td><td>14</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0927" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/27</h3></span>
 +
<span class="keln_exp"><h4>Reverse Transcription</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>number</th><th>template/&microl</th><th>RTase/&microl</th><th>RNAase free water/&microl</th><th>RT buffer/&microl</th><th>Primer Mix/&microl</th><th>Total</th></tr>
 +
<tr><td>A1</td><td>BclAcex/14</td><td>1</td><td>0</td><td>4</td><td>1</td><td>20</td></tr>
 +
<tr><td>A2</td><td>BclAclos/14</td><td>1</td><td>0</td><td>4</td><td>1</td><td>20</td></tr>
 +
<tr><td>A3</td><td>INPcex/14</td><td>1</td><td>0</td><td>4</td><td>1</td><td>20</td></tr>
 +
<tr><td>A4</td><td>INPclos/14</td><td>1</td><td>0</td><td>4</td><td>1</td><td>20</td></tr>
 +
<tr><td>A5</td><td>BclAscFv/14</td><td>1</td><td>0</td><td>4</td><td>1</td><td>20</td></tr>
 +
<tr><td>A6</td><td>INPscFv/12</td><td>1</td><td>0</td><td>4</td><td>1</td><td>20</td></tr>
 +
<tr><td>B1</td><td>BclAcex/14</td><td>0</td><td>1</td><td>4</td><td>1</td><td>20</td></tr>
 +
<tr><td>B2</td><td>BclAclos/14</td><td>0</td><td>1</td><td>4</td><td>1</td><td>20</td></tr>
 +
<tr><td>B3</td><td>INPcex/14</td><td>0</td><td>1</td><td>4</td><td>1</td><td>20</td></tr>
 +
<tr><td>B4</td><td>INPclos/14</td><td>0</td><td>1</td><td>4</td><td>1</td><td>20</td></tr>
 +
<tr><td>B5</td><td>BclAscFv/14</td><td>0</td><td>1</td><td>4</td><td>1</td><td>20</td></tr>
 +
<tr><td>B6</td><td>INPscFv/12</td><td>0</td><td>1</td><td>4</td><td>1</td><td>20</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0927" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/27</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder/5</td><td></td></tr>
 +
<tr><td>2</td><td>9/26 PCR INP- His scFv/25</td><td></td></tr>
 +
<tr><td>3</td><td>9/26 PCR INP- His scFv/25</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/e/ed/T--Kyoto--9_27_No152_Electrophoresis_Miyazaki_10_06_04_10_12.1.jpg"><img src="https://static.igem.org/mediawiki/2016/a/a0/T--Kyoto--9_27_No152_Electrophoresis_Miyazaki_10_06_04_10_12.2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0927" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/27</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>9/27 electrophoresis lane2,3</td><td></td><td>0.7660g</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">nanodrop<br>17.8ng/nl<br>260/280 1.79<br>260/230 0.31</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0927" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/27</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>15th</th><th>null</th><th>null</th><th>null</th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>BclAcex+ (CB buffer)</td><td>0.181/0.212/0.204</td><td>0.254/0.255/0.254</td><td>0/0/0</td></tr>
 +
<tr><td>BclAcex- (CB buffer)</td><td>0.208/0.275/0.228</td><td>0.160/0.171/0.162</td><td></td></tr>
 +
<tr><td>BclActl+ (CB buffer)</td><td>0.178/0.176/0.171</td><td>0.082/0.085/0.082</td><td>0.022/0.02/0.02</td></tr>
 +
<tr><td>BclActl- (CB buffer)</td><td>0.218/0.214/0.216</td><td>0.161/0.160/0.159</td><td></td></tr>
 +
<tr><td>BclAcex+ (PBS buffer)</td><td>0.212/0.211/0.210</td><td>0.333/0.334/0.333</td><td>0.022/0.022/0.021</td></tr>
 +
<tr><td>BclAcex- (PBS buffer)</td><td>0.203/0.202/0.201</td><td>0.178/0.178/0.180</td><td></td></tr>
 +
<tr><td>BclActl+ (PBS buffer)</td><td>0.182/0.181/0.180</td><td>0.253/0.246/0.246</td><td>0.021/0.021/0.021</td></tr>
 +
<tr><td>BclActl- (PBS buffer)</td><td>0.208/0.205/0.208</td><td>0.192/0.194/0.194</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0929" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/29</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>9/27 Gel Extract/2</td><td>VF/0.75</td><td>VR/0.75</td><td>2×KAPA HiFi Hotstart ReadyMix/12.5</td><td>9</td><td></td><td>25</td></tr>
 +
<tr><td></td><td>VF/0.75</td><td>VR/0.75</td><td>2×KAPA HiFi Hotstart ReadyMix/12.5</td><td>9</td><td></td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0929" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/29</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>BAN scFv</td><td>LB</td></tr>
 +
<tr><td>INP clos</td><td>LB</td></tr>
 +
<tr><td>BAN clos</td><td>LB</td></tr>
 +
<tr><td>INP cex</td><td>LB</td></tr>
 +
<tr><td>BAN cex</td><td>LB</td></tr>
 +
<tr><td>K875004</td><td>LB</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0929" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/29</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder/2</td><td></td></tr>
 +
<tr><td>2</td><td>9/29 PCR INP-His-scFv/2</td><td></td></tr>
 +
<tr><td>3</td><td></td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/3/3e/T--Kyoto--9_29_No157_Electrophoresis_Miyazaki_10_06_04_22_37.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0929" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/29</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>9/29 PCR INP-His-scFv</td><td>31.9</td><td>2.02</td><td>1.38</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="0930" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/30</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino, Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td></td></tr>
 +
<tr><td>2</td><td>BclA clos/4.0</td><td></td></tr>
 +
<tr><td>3</td><td>INP clos/5.5</td><td></td></tr>
 +
<tr><td>4</td><td>BclA cex/4.5</td><td></td></tr>
 +
<tr><td>5</td><td>INP cex/2.5</td><td></td></tr>
 +
<tr><td>6</td><td>BclA scFv/4.5</td><td></td></tr>
 +
<tr><td>7</td><td>INP scFv/4.0</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0905" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/5</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>8/10 J23105 RFP miniprep </td><td>20</td><td></td><td>1</td><td>0</td><td>1</td><td>10×H 3</td><td>3</td><td>2</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0905" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/5</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>8/29 Ligation Bclascfv-BclaCBDcex</td><td>2</td><td>CH5a/20</td><td>LB(CP)</td></tr>
 +
<tr><td>〃Bclascfv-BclaCBDclos</td><td>2</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃Bclascfv-INPCBDcex</td><td>2</td><td>〃</td><td>〃</td></tr>
 +
<tr><td>〃Bclascfv-INPCBDclos</td><td>2</td><td>〃</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0905" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/5</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>8/20 SPcut Bcla-CBDcex/25</td><td>2520</td></tr>
 +
<tr><td>2</td><td>〃Bcla-CBDclos/25</td><td>〃</td></tr>
 +
<tr><td>3</td><td>〃INP-CBDcex/25</td><td>〃</td></tr>
 +
<tr><td>4</td><td>〃INP-CBDclos/25</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/8/8a/T--Kyoto--9_5_No74_Electrophoresis_Yamamoto_09_30_10_51_18.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Kosaka -->
 +
<div class="keln_container">
 +
<a name="0905" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>9/5</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>100bp ladder/5</td><td>6×loading Dye -</td></tr>
 +
<tr><td>2</td><td>8/10 miniprep 9/5 xpcut J23105RFP/30</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1001" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/1</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>3RT- target /10</td><td></td></tr>
 +
<tr><td>2</td><td>3RT+ target /10</td><td></td></tr>
 +
<tr><td>3</td><td>3RT- 16s rRNA /10</td><td></td></tr>
 +
<tr><td>4</td><td>3RT+ 16s rRNA /10</td><td></td></tr>
 +
<tr><td>5</td><td>4RT- target /10</td><td></td></tr>
 +
<tr><td>6</td><td>4RT+ target /10</td><td></td></tr>
 +
<tr><td>7</td><td>4RT- 16s rRNA /10</td><td></td></tr>
 +
<tr><td>8</td><td>4RT+ 16s rRNA /10</td><td></td></tr>
 +
<tr><td>9</td><td>100bp ladder /5</td><td></td></tr>
 +
<tr><td>10</td><td>A-6 target /10</td><td></td></tr>
 +
<tr><td>11</td><td>A-5 target /10</td><td></td></tr>
 +
<tr><td>12</td><td>A-6 16s rRNA /10</td><td></td></tr>
 +
<tr><td>13</td><td>A-5 16s rRNA /10</td><td></td></tr>
 +
<tr><td>14</td><td>B-6 target /10</td><td></td></tr>
 +
<tr><td>15</td><td>B-5 target /10</td><td></td></tr>
 +
<tr><td>16</td><td>B-6 16s rRNA /10</td><td></td></tr>
 +
<tr><td>17</td><td>B-5 16s rRNA /10</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/4/4a/T--Kyoto--10_1_No165_Electrophoresis_Miyazaki_10_10_07_19_39.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1001" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/1</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>K165002</td><td></td><td>DH5α</td><td></td></tr>
 +
<tr><td>BclA scFv</td><td></td><td>DH5α</td><td></td></tr>
 +
<tr><td>INP scFv</td><td></td><td>DH5α</td><td></td></tr>
 +
<tr><td>BclA cex</td><td></td><td>DH5α</td><td></td></tr>
 +
<tr><td>INP cex</td><td></td><td>DH5α</td><td></td></tr>
 +
<tr><td>BclA clos</td><td></td><td>DH5α</td><td></td></tr>
 +
<tr><td>INP clos</td><td></td><td>DH5α</td><td></td></tr>
 +
<tr><td>K165002</td><td></td><td>BL21</td><td></td></tr>
 +
<tr><td>BclA scFv</td><td></td><td>BL21</td><td></td></tr>
 +
<tr><td>INP scFv</td><td></td><td>BL21</td><td></td></tr>
 +
<tr><td>BclA cex</td><td></td><td>BL21</td><td></td></tr>
 +
<tr><td>INP cex</td><td></td><td>BL21</td><td></td></tr>
 +
<tr><td>BclA clos</td><td></td><td>BL21</td><td></td></tr>
 +
<tr><td>INP clos</td><td></td><td>BL21</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1004" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/4</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>INP CTL 8/4 miniprep</td><td></td><td>DH5α/20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1005" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/5</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>INP CTL 10/4 transformation</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1005" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/5</h3></span>
 +
<span class="keln_exp"><h4>Western Blotting</h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>lane</th><th>sample</th></tr>
 +
<tr><td>Marker</td><td>M.Marker(Wako) 5&microl</td></tr>
 +
<tr><td>1</td><td>BBa_E165002(NC) </td></tr>
 +
<tr><td>2</td><td>BclA-6His-scFv  (33 kDa) </td></tr>
 +
<tr><td>3</td><td>INPNC-6His-scFv  (63 kDa) </td></tr>
 +
<tr><td>4</td><td>BclA-6His-CBDcex (17 kDa) </td></tr>
 +
<tr><td>5</td><td>INPNC-6His-CBDcex(47kDa) </td></tr>
 +
<tr><td>6</td><td>BclA-6His-CBDclos (17 kDa) </td></tr>
 +
<tr><td>7</td><td>INPNC-6His-CBDclos (47 kDa) </td></tr>
 +
<tr><td>8</td><td>BBa_K875004(PC) Lpp-OmpA-scFv-6His (43 kDa)</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/c/cb/T--Kyoto--10_5_No193_General_Miyazaki_10_17_07_20_04.png">
 +
</div>
 +
<p class="keln_note">Used pelet from 400 &microl overnight culture.<br><br>Transfar ;80 min 200 mA(CC)<br>Blocking ;O/N<br>1’antibody ;anti-6His(mouse)<br>1:1000 with blocking buffer 1h<br>2’antibody ;anti-mouse HRP　1:2500 with TBST 1h<br>Detection time　240 sec<br>(ECL Prime)</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1006" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/6</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>10/5 liquid culture INP CTL /2.5ml</td><td>LB(CP) /47.5ml</td></tr>
 +
<tr><td>10/5 liquid culture INP CTL /2.5ml</td><td>LB(CP) /47.5ml</td></tr>
 +
<tr><td>10/5 liquid culture INP CBD cex(5) /2.5ml</td><td>LB(CP) /47.5ml</td></tr>
 +
<tr><td>10/5 liquid culture INP CBD cex(5) /2.5ml</td><td>LB(CP) /47.5ml</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1006" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/6</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>10/5 liquid culture INP CTL /100ml</td><td>LB(CP) /100ml</td></tr>
 +
<tr><td>10/3 liquid culture INP CBD cex /100ml</td><td>LB(CP) /200ml</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1007" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/7</h3></span>
 +
<span class="keln_exp"><h4>CBD</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>16th</th><th>null</th><th>null</th><th>null</th></tr>
 +
<tr><td>sample</td><td>before</td><td>after</td><td>wash</td></tr>
 +
<tr><td>INPcex+</td><td>0.208/0.208/0.211</td><td>0.224/0.221/0.226</td><td>0.013/0.012/0.010</td></tr>
 +
<tr><td>INPcex-</td><td>0.210/0.216/0.204</td><td>0.232/0.231/0.232</td><td></td></tr>
 +
<tr><td>INPctl+</td><td>0.196/0.196/0.191</td><td>0.220/0.217/0.212</td><td>0.023/0.024/0.025</td></tr>
 +
<tr><td>INPctl-</td><td>0.194/0.195/0.196</td><td>0.207/0.199/0.208</td><td></td></tr>
 +
<tr><td>cellulose only</td><td>0.015/0.013/0.015</td><td>0.031/0.031/0.032</td><td>0.018/0.017/0.019</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1011" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/11</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>2/25 miniprep INPcex</td><td>3</td><td>DH5α /20</td><td>LB(CP)×3</td></tr>
 +
<tr><td>2/8 miniprep INP His 12 (INPscFv)</td><td>3</td><td>DH5α /20</td><td>LB(CP)×3</td></tr>
 +
<tr><td>2/25 miniprep INPcex</td><td>3</td><td>BL21 /20</td><td>LB(CP)×3</td></tr>
 +
<tr><td>2/8 miniprep INP His 12 (INPscFv)</td><td>3</td><td>BL21 /20</td><td>LB(CP)×3</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1011" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/11</h3></span>
 +
<span class="keln_exp"><h4>SEM(CBD)</h4></span>
 +
<span class="keln_researcher">Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>sample/ml</th><th>before</th><th>after</th><th>wash1</th><th>wash2</th></tr>
 +
<tr><td>INP CBDcex 10/1 liquid culture /90</td><td>0.550</td><td>0.392</td><td>0.074/0.084</td><td>-0.004/-0.018</td></tr>
 +
<tr><td>INP CBDcex 10/1 liquid culture /45</td><td>0.550</td><td>0.432</td><td>0.002/0.000</td><td>-0.001/0.000</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1012" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/12</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony1</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony2</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony3</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony4</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony5</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony6</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony7</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony8</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony9</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony10</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony11</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony12</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony13</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony14</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony15</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony16</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony17</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony18</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony19</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony20</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1012" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/12</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony21</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony21</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony21</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>INP-His-scFv-INP-His-CBDcex DH5α colony21</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>2/8 miniprep INP-His-scFv /0.5</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
<tr><td>2/25 miniprep INP-His-CBDcex /0.5</td><td>VF/0.5 VR/0.5</td><td>11.5</td><td>Quick Taq/12.5</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1012" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/12</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony1 /5</td><td></td></tr>
 +
<tr><td>2</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony2 /5</td><td></td></tr>
 +
<tr><td>3</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony3 /5</td><td></td></tr>
 +
<tr><td>4</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony4 /5</td><td></td></tr>
 +
<tr><td>5</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony5 /5</td><td></td></tr>
 +
<tr><td>6</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony6 /5</td><td></td></tr>
 +
<tr><td>7</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony7 /5</td><td></td></tr>
 +
<tr><td>8</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony8 /5</td><td></td></tr>
 +
<tr><td>9</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony17 /5</td><td></td></tr>
 +
<tr><td>10</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony18 /5</td><td></td></tr>
 +
<tr><td>11</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony19 /5</td><td></td></tr>
 +
<tr><td>12</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony20 /5</td><td></td></tr>
 +
<tr><td>13</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony21 /5</td><td></td></tr>
 +
<tr><td>14</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony22 /5</td><td></td></tr>
 +
<tr><td>15</td><td>INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony23 /5</td><td></td></tr>
 +
<tr><td>16</td><td>INPNC-His-CBDcex 10/12 colony PCR /5</td><td></td></tr>
 +
<tr><td>17</td><td>1kbp ladder /10</td><td></td></tr>
 +
<tr><td></td><td> </td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/1/11/T--Kyoto--10_12_No172_Electrophoresis_Miyazaki_10_15_05_41_02.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1012" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/12</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/8 INP-His 12 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/8 INP-His 12 /0.1</td><td>VF /0.75</td><td>VR /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>sample /&microl</th><th>primer /&microl</th><th>KAPA Taq Extra Hotstart /&microl</th><th>DMSO /&microl</th><th>MilliQ /&microl</th><th>total /&microl</th></tr>
 +
<tr><td>INP-cex + INP-scFv colony2</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony7</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony17</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony19</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony9</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony10</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony11</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony12</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony13</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony16</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony23</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony24</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony25</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony26</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony27</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony28</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony29</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony30</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony31</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>sample /&microl</th><th>primer /&microl</th><th>KAPA Taq Extra Hotstart /&microl</th><th>DMSO /&microl</th><th>MilliQ /&microl</th><th>total /&microl</th></tr>
 +
<tr><td>INP-cex + INP-scFv colony31</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony32</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony33</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex + INP-scFv colony34</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-scFv /0.25</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
<tr><td>INP-cex /0.25</td><td>VF /1.25  VR /1.25</td><td>12.5</td><td>0.75</td><td>9.25</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>10/6 liquid culture INP-His-CBDcex</td><td>27.8</td><td>1.96</td><td>1.86</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>10/12 PCR INP-His 12 (INP-His coding)</td><td>727.3</td><td>1.88</td><td>1.86</td></tr>
 +
<tr><td>10/13 PCR INP-His 12</td><td>1132.1</td><td>1.89</td><td>2.04</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>10/13 PCR purification INP-His coding</td><td>7</td><td>2</td><td>0</td><td>0</td><td>2</td><td>10×H /6</td><td>0</td><td>44</td><td>60</td></tr>
 +
<tr><td>10/13 PCR purification INP-His 12</td><td>6</td><td>0</td><td>2</td><td>0</td><td>2</td><td>10×M /6</td><td>0</td><td>44</td><td>60</td></tr>
 +
<tr><td>10/13 midiprep INP-His-CBDcex</td><td>50</td><td>2</td><td>0</td><td>0</td><td>2</td><td>10×H /6</td><td>0</td><td>0</td><td>60</td></tr>
 +
<tr><td>10/13 midiprep INP-His-CBDcex</td><td>50</td><td>0</td><td>0</td><td>2</td><td>2</td><td>10×H /6</td><td>0</td><td>0</td><td>60</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>INPcex+INPscFv colony2</td><td></td></tr>
 +
<tr><td>2</td><td>INPcex+INPscFv colony7</td><td></td></tr>
 +
<tr><td>3</td><td>INPcex+INPscFv colony19</td><td></td></tr>
 +
<tr><td>4</td><td>INPcex+INPscFv colony9</td><td></td></tr>
 +
<tr><td>5</td><td>INPcex+INPscFv colony17</td><td></td></tr>
 +
<tr><td>6</td><td>INPcex+INPscFv colony10</td><td></td></tr>
 +
<tr><td>7</td><td>INPcex+INPscFv colony11</td><td></td></tr>
 +
<tr><td>8</td><td>INPcex+INPscFv colony12</td><td></td></tr>
 +
<tr><td>9</td><td>INPcex+INPscFv colony15</td><td></td></tr>
 +
<tr><td>10</td><td>INPcex+INPscFv colony16</td><td></td></tr>
 +
<tr><td>11</td><td>INPcex+INPscFv colony23</td><td></td></tr>
 +
<tr><td>12</td><td>INPcex+INPscFv colony24</td><td></td></tr>
 +
<tr><td>13</td><td>INPcex+INPscFv colony25</td><td></td></tr>
 +
<tr><td>14</td><td>INPcex+INPscFv colony26</td><td></td></tr>
 +
<tr><td>15</td><td>INPscFv /5</td><td></td></tr>
 +
<tr><td>16</td><td>INPcex /5</td><td></td></tr>
 +
<tr><td>17</td><td>1kbp ladder</td><td></td></tr>
 +
</table>
 +
<img src="10_13_No178_Electrophoresis_Miyazaki_10_16_03_04_04.jpg
 +
https://static.igem.org/mediawiki/2016/2/26/T--Kyoto--">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>A1</td><td>INPcex+INPscFv colony2</td><td></td></tr>
 +
<tr><td>A2</td><td>INPcex+INPscFv colony7</td><td></td></tr>
 +
<tr><td>A3</td><td>INPcex+INPscFv colony19</td><td></td></tr>
 +
<tr><td>A4</td><td>INPcex+INPscFv colony9</td><td></td></tr>
 +
<tr><td>A5</td><td>INPcex+INPscFv colony17</td><td></td></tr>
 +
<tr><td>A6</td><td>INPscFv /5</td><td></td></tr>
 +
<tr><td>A7</td><td>INPcex /5</td><td></td></tr>
 +
<tr><td>A8</td><td>1kbp ladder /5</td><td></td></tr>
 +
<tr><td>B1</td><td>INPcex+INPscFv colony10</td><td></td></tr>
 +
<tr><td>B2</td><td>INPcex+INPscFv colony11</td><td></td></tr>
 +
<tr><td>B3</td><td>INPcex+INPscFv colony12</td><td></td></tr>
 +
<tr><td>B4</td><td>INPcex+INPscFv colony13</td><td></td></tr>
 +
<tr><td>B5</td><td>INPcex+INPscFv colony14</td><td></td></tr>
 +
<tr><td>B6</td><td>1kbp ladder /5</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/2/26/T--Kyoto--10_13_No179_Electrophoresis_Miyazaki_10_16_03_20_40.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kbp ladder</td><td></td></tr>
 +
<tr><td>2</td><td>10/13 1A</td><td></td></tr>
 +
<tr><td>3</td><td>10/13 2A</td><td></td></tr>
 +
<tr><td>4</td><td>10/13 3A</td><td></td></tr>
 +
<tr><td>5</td><td>10/13 4A</td><td></td></tr>
 +
<tr><td>6</td><td>10/13 5A</td><td></td></tr>
 +
<tr><td>7</td><td>10/13 1B</td><td></td></tr>
 +
<tr><td>8</td><td>10/13 2B</td><td></td></tr>
 +
<tr><td>9</td><td>10/13 3B</td><td></td></tr>
 +
<tr><td>10</td><td>10/13 4B</td><td></td></tr>
 +
<tr><td>11</td><td>10/15 5B</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/e/e2/T--Kyoto--10_13_No180_Electrophoresis_Miyazaki_10_16_03_25_22.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>1A+2A</td><td>10/13 PCR purification INP His coding</td><td></td></tr>
 +
<tr><td>3A~5A</td><td>10/13 midiprep INP His CBDcex</td><td></td></tr>
 +
<tr><td>1B+2B</td><td>10/13 PCR purification INP His12</td><td></td></tr>
 +
<tr><td>3B~5B</td><td>10/13 midiprep INP His CBDcex</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/0/05/T--Kyoto--10_13_No181_Gel_Extraction_Miyazaki_10_16_03_33_03.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>Fluorescence Microscopy</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>sample</th><th>number</th><th>magnification</th></tr>
 +
<tr><td>INPclos</td><td>13318</td><td>×200</td></tr>
 +
<tr><td>INPclos</td><td>13319~13328</td><td>×600</td></tr>
 +
<tr><td>INP CTL</td><td>13329</td><td>×200</td></tr>
 +
<tr><td>INP CTL</td><td>13330~13338</td><td>×600</td></tr>
 +
<tr><td>cellulose only</td><td>13339</td><td>×200</td></tr>
 +
<tr><td>cellulose only</td><td>13340,13343~13351</td><td>×600</td></tr>
 +
<tr><td>cellulose only</td><td>13341,13342</td><td>×1000</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">without filter</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>Fluorescence Microscopy</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>E.coli</th><th>OD600</th><th>cellulose</th><th>weight</th><th>filter</th></tr>
 +
<tr><td>-</td><td>-</td><td>powder</td><td>10</td><td>non-filtered</td></tr>
 +
<tr><td>INPNC-His-ctl</td><td>0.2</td><td>powder</td><td>10</td><td>non-filtered</td></tr>
 +
<tr><td>INPNC-His-CBDcex</td><td>0.2</td><td>powder</td><td>10</td><td>non-filtered</td></tr>
 +
<tr><td>-</td><td>-</td><td>powder</td><td>10</td><td>filtered</td></tr>
 +
<tr><td>INPNC-His-ctl</td><td>0.2</td><td>powder</td><td>10</td><td>filtered</td></tr>
 +
<tr><td>INPNC-His-CBDcex</td><td>0.2</td><td>powder</td><td>10</td><td>filtered</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>Fluorescence Microscopy</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>sample</th><th>number</th><th>magnification</th></tr>
 +
<tr><td>INPcex</td><td>13353</td><td>×200</td></tr>
 +
<tr><td>INPcex</td><td>13354~13356,13364,13366</td><td>×600</td></tr>
 +
<tr><td>INPcex</td><td>13357,13365</td><td>×1000</td></tr>
 +
<tr><td>INP CTL</td><td>13367</td><td>×200</td></tr>
 +
<tr><td>INP CTL</td><td>13368~13378</td><td>×600</td></tr>
 +
<tr><td>INP CTL</td><td>13379</td><td>×1000</td></tr>
 +
<tr><td>cellulose only</td><td>13380,13381</td><td>×200</td></tr>
 +
<tr><td>cellulose only</td><td>13382~13391</td><td>×600</td></tr>
 +
<tr><td>cellulose only</td><td></td><td></td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">with filter</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>Fluorescence Microscopy</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>sample</th><th>number</th><th>magnification</th></tr>
 +
<tr><td>INPcex</td><td>13392~98</td><td>×200</td></tr>
 +
<tr><td>INPcex</td><td>13399~13408</td><td>×600</td></tr>
 +
<tr><td>INP CTL</td><td>13309~13419</td><td>×200</td></tr>
 +
<tr><td>bemliese</td><td>13420~13430</td><td>×200</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">with bemliese</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1014" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/14</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kbp ladder /3</td><td></td></tr>
 +
<tr><td>2</td><td>10/13 1A /25</td><td></td></tr>
 +
<tr><td>4</td><td>10/13 2A /25</td><td></td></tr>
 +
<tr><td>5</td><td>10/13 3A /25</td><td></td></tr>
 +
<tr><td>6</td><td>10/13 4A /25</td><td></td></tr>
 +
<tr><td>7</td><td>10/13 5A /25</td><td></td></tr>
 +
<tr><td>8</td><td>10/13 1B /25</td><td></td></tr>
 +
<tr><td>9</td><td>10/13 2B /25</td><td></td></tr>
 +
<tr><td>10</td><td>10/13 3B /25</td><td></td></tr>
 +
<tr><td>11</td><td>10/13 4B /25</td><td></td></tr>
 +
<tr><td>12</td><td>10/13 5B /25</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/b/b9/T--Kyoto--10_14_No186_Electrophoresis_Miyazaki_10_16_04_07_57.1.jpg"><img src="https://static.igem.org/mediawiki/2016/8/8a/T--Kyoto--10_14_No186_Electrophoresis_Miyazaki_10_16_04_07_57.2.jpg">
 +
</div>
 +
<p class="keln_note">with bemliese</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1014" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/14</h3></span>
 +
<span class="keln_exp"><h4>SEM(CBD)</h4></span>
 +
<span class="keln_researcher">Miyazaki</span>
 +
<table class="keln_table">
 +
<tr><th>sample</th><th>membrane shape</th></tr>
 +
<tr><td>INPcex + cellulose</td><td>square</td></tr>
 +
<tr><td>INP CTL + cellulose</td><td>triangle</td></tr>
 +
<tr><td>cellulose only</td><td>pentagon</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1014" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/14</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>10/14 gel extract e2 /5</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/a/a0/T--Kyoto--10_14_No188_Electrophoresis_Miyazaki_10_16_06_42_15.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1014" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/14</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>10/14 electrophoresis 1B longer + 2B longer e1</td><td>10/13 restriction enzyme design INP His 12</td><td></td></tr>
 +
<tr><td>10/14 electrophoresis 1B longer + 2B longer e2</td><td>10/13 restriction enzyme design INP His12</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1014" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/14</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kbp ladder /3</td><td></td></tr>
 +
<tr><td>2</td><td>1A + 2A /3</td><td></td></tr>
 +
<tr><td>3</td><td>3A + 4A + 5A /3</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/7/7c/T--Kyoto--10_14_No189_Electrophoresis_Miyazaki_10_16_07_13_08.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1014" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/14</h3></span>
 +
<span class="keln_exp"><h4>Gel Extraction</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>Restriction Enzyme Digestion Product</th><th>Volume/(&microl)</th></tr>
 +
<tr><td>10/14 electrophoresis 1B + 2B</td><td>10/13 restriction enzyme design INP His12</td><td></td></tr>
 +
<tr><td>10/14 electrophoresis 1A + 2A</td><td>10/13 restriction enzyme design INP His coding</td><td></td></tr>
 +
<tr><td>10/14 electrophoresis 3A + 4A + 5A</td><td>INP His CBDcex</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1014" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/14</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>sample1 1A + 2A /&microl</th><th>sample2 3A~5A /&microl</th><th>Ligation high ver.2</th><th>total</th></tr>
 +
<tr><td>5</td><td>2.5</td><td>7.5</td><td>15</td></tr>
 +
<tr><td>3.75</td><td>3.75</td><td>7.5</td><td>15</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1014" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/14</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>INP His coding ligationA</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>INP His coding ligationA</td><td>4</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>INP His coding ligationB</td><td>2</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
<tr><td>INP His coding ligationB</td><td>4</td><td>DH5α /20</td><td>LB(CP)</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1015" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/15</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher"></span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>10/15 INP-His coding i</td><td>207.4</td><td>1.87</td><td>2.11</td></tr>
 +
<tr><td>10/15 INP-His coding G</td><td>174.7</td><td>1.88</td><td>1.09</td></tr>
 +
<tr><td>10/15 INP-His coding E</td><td>175.5</td><td>1.08</td><td>2.10</td></tr>
 +
<tr><td>10/15 INP-His coding M</td><td>212.9</td><td>1.88</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1015" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/15</h3></span>
 +
<span class="keln_exp"><h4>CBD　Binding Assay</h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>sample</th><th>Buffer</th><th>OD</th></tr>
 +
<tr><td>INP cex</td><td>CBC buffer</td><td>1.002</td></tr>
 +
<tr><td>simultaneous expression</td><td>CBC buffer</td><td>0.994</td></tr>
 +
<tr><td>INP CTL</td><td>CBC buffer</td><td>1.004</td></tr>
 +
<tr><td>INP CTL</td><td>PBS</td><td>0.964</td></tr>
 +
<tr><td>simultaneous expression</td><td>PBS</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1015" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/15</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
 +
<span class="keln_researcher"></span>
 +
<table class="keln_table">
 +
<tr><th>Sample</th><th>DNA/(&microl)</th><th>EcoR1/(&microl)</th><th>Xba1/(&microl)</th><th>Spe1/(&microl)</th><th>Pst1/(&microl)</th><th>Buffer/(&microl)</th><th>BSA/(&microl)</th><th>MilliQ/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>10/15 PCR purification INP His coding i</td><td>29.0</td><td>2</td><td>0</td><td>0</td><td>2</td><td>6</td><td></td><td>21</td><td>60</td></tr>
 +
<tr><td>10/15 PCR purification INP His coding G</td><td>34.3</td><td>2</td><td>0</td><td>0</td><td>2</td><td>6</td><td></td><td>15.7</td><td>60</td></tr>
 +
<tr><td>10/15 PCR purification INP His coding E</td><td>34.2</td><td>2</td><td>0</td><td>0</td><td>2</td><td>6</td><td></td><td>15.8</td><td>60</td></tr>
 +
<tr><td>10/15 PCR purification INP His coding M</td><td>28.2(6000ng)</td><td>2</td><td>0</td><td>0</td><td>2</td><td>6</td><td></td><td>21.8</td><td>60</td></tr>
 +
<tr><td>10/13 midiprep INP-His-CBDcex</td><td>50</td><td>2</td><td>0</td><td>0</td><td>2</td><td>6</td><td></td><td>0</td><td>60</td></tr>
 +
<tr><td>10/13 midiprep INP-His-CBDcex</td><td>50</td><td>2</td><td>0</td><td>0</td><td>2</td><td>6</td><td></td><td>0</td><td>60</td></tr>
 +
<tr><td>10/13 midiprep INP-His-CBDcex</td><td>50</td><td>2</td><td>0</td><td>0</td><td>2</td><td>6</td><td></td><td>0</td><td>60</td></tr>
 +
<tr><td>10/13 midiprep INP-His-CBDcex</td><td>50</td><td>2</td><td>0</td><td>0</td><td>2</td><td></td><td></td><td>0</td><td>60</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">1.5　Dr.Kitabatake<br>2.6 Agarose S ＋　Promega Gel Extraction<br>3.7 Agarose S ＋　Dr.Nasuda<br>4.8　stock</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1015" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/15</h3></span>
 +
<span class="keln_exp"><h4>Gel strecking</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>sample</th><th>null</th></tr>
 +
<tr><td>INPNc-His-scFv </td><td>(CP)(LB)plate×3</td></tr>
 +
<tr><td>INP-His-CBDcex</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1015" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/15</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
<tr><td>2/22 INP-His scFv colony3 /0.1</td><td>INP His coding Fw /0.75</td><td>INP His coding Rv /0.75</td><td>2×KAPA /12.5</td><td>10.9</td><td>0</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1015" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/15</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>A1 10/14 INPcex + INPscFv colony1</td><td>VF/1.25 VR/1.25</td><td>8.75</td><td>KAPA2G /12.5 DMSO/1.25</td><td>25</td></tr>
 +
<tr><td>A2 10/14 INPcex + INPscFv colony2</td><td>VF/1.25 VR/1.25</td><td>8.75</td><td>KAPA2G /12.5 DMSO/1.25</td><td>25</td></tr>
 +
<tr><td>A3 10/14 INPcex + INPscFv colony3</td><td>VF/1.25 VR/1.25</td><td>8.75</td><td>KAPA2G /12.5 DMSO/1.25</td><td>25</td></tr>
 +
<tr><td>A4 10/14 INPcex + INPscFv colony4</td><td>VF/1.25 VR/1.25</td><td>8.75</td><td>KAPA2G /12.5 DMSO/1.25</td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1015" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/15</h3></span>
 +
<span class="keln_exp"><h4>Western Blotting</h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>lane</th><th>sample</th></tr>
 +
<tr><td>Marker</td><td>M.Marker</td></tr>
 +
<tr><td>1</td><td>BBa_K875004(PC)    Lpp-OmpA-scFv-6His 43kDa </td></tr>
 +
<tr><td>2</td><td>INPNC-6His-CBDcex 47kDa </td></tr>
 +
<tr><td>3</td><td>INPNC-6His-scFv      63kDa </td></tr>
 +
<tr><td>4</td><td>INPNC-6His-CBDcex/INPNC-6His-scFv</td></tr>
 +
<tr><td>5</td><td>BBa_K165002(NC) </td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/0/03/T--Kyoto--10_15_No194_General_Miyazaki_10_17_08_24_33.png">
 +
</div>
 +
<p class="keln_note">Used pelet from 500 &microl overnight culture, adjested OD600 to 3.1 .<br><br>Transfar ;80 min 200 mA(CC)<br>Blocking ;O/N<br>1’antibody ;anti-6His(mouse)<br>1:1000 with blocking buffer 1h<br>2’antibody ;anti-mouse HRP　1:2500 with TBST 1h<br>Detection time　900 sec<br>(ECL Prime)</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1kb ladder</td><td></td><td></td></tr>
 +
<tr><td>10/15 colony PCR colony 1</td><td>3</td><td></td></tr>
 +
<tr><td>10/15 colony PCR colony 2</td><td>25</td><td></td></tr>
 +
<tr><td>10/15 colony PCR colony 3</td><td>25</td><td></td></tr>
 +
<tr><td>10/15 colony PCR colony 4</td><td></td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>Cellulose binding Assay</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Cellulose/(mg)</th><th>E.coli/(mg)</th><th>buffer</th></tr>
 +
<tr><td>1</td><td>cex/1</td><td>carbonate-bicarbonate buffer</td></tr>
 +
<tr><td>1</td><td>ctl/1</td><td>carbonate-bicarbonate buffer</td></tr>
 +
<tr><td>0.5</td><td>cex/1</td><td>carbonate-bicarbonate buffer</td></tr>
 +
<tr><td>0.5</td><td>ctl/1</td><td>carbonate-bicarbonate buffer</td></tr>
 +
<tr><td>0.5</td><td>CBC/1</td><td>carbonate-bicarbonate buffer</td></tr>
 +
<tr><td>0.1</td><td>cex/1</td><td>carbonate-bicarbonate buffer</td></tr>
 +
<tr><td>0.1</td><td>ctl/1</td><td></td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">RT,-3h 500rpm 15:45~21:15 binding<br>fix with 4×1/100 PFA 10min→centrifuge→PBS→centrifuge<br>→PBS→centrifuge→PBS→centrifuge→DAP1→centrifuge<br>→PBS→centrifuge→PBS→through filter→PBS into filter<br>→centrifuge→PBSinto filter→centrifuge<br>→suspend with PBS→centrifuge<br>→suspend with?discoloration inhibitor<br></p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>Dual Display Assay for Electron Microscope</h4></span>
 +
<span class="keln_researcher"></span>
 +
<table class="keln_table">
 +
<tr><th>E.coli</th><th>Cellulose</th><th>VLP</th><th>buffer</th></tr>
 +
<tr><td>A.Dual 1mL</td><td>1mg</td><td>10&microL</td><td>PBS</td></tr>
 +
<tr><td>B.Dual 1mL</td><td>1mg</td><td></td><td>PBS</td></tr>
 +
<tr><td>C.Dual 1mL</td><td></td><td>10&microL</td><td>PBS</td></tr>
 +
<tr><td>D.Dual 1mL</td><td></td><td></td><td>PBS</td></tr>
 +
<tr><td>E.INPctl</td><td>1mg</td><td>10&microL</td><td>PBS</td></tr>
 +
<tr><td>F.INPctl</td><td>1mg</td><td></td><td>PBS</td></tr>
 +
<tr><td>G.INPctl</td><td></td><td>10&microL</td><td>PBS</td></tr>
 +
<tr><td>H.INPctl</td><td></td><td></td><td>PBS</td></tr>
 +
<tr><td>I</td><td>1mg</td><td></td><td>PBS</td></tr>
 +
<tr><td>J</td><td></td><td>10&microL</td><td></td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">RT,ih incubate,850rpm<br>Dul=1.020<br>ctl=1.056</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher"></span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1 kb ladder</td><td>5</td></tr>
 +
<tr><td>2</td><td>10/15 Digestion INP His Coding 2</td><td>30 /0.94 上0.04下0.10</td></tr>
 +
<tr><td>3</td><td>10/15 Digestion INP His Coding 2</td><td>30  /0.94 上0.04下0.19</td></tr>
 +
<tr><td>4</td><td>10/13 miniprep INP-His-CBDcex 6</td><td>30 /0.94 0.11</td></tr>
 +
<tr><td>5</td><td>10/13 miniprep INP-His-CBDcex 6</td><td>30 /0.94</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">Uchino 16:08~<br>ABEF→centrifuge→resupend with PBS →centrifuge→through filter→resupend with PBS→through filter→centrifuge→respend with PBS→centrifuge→resupend with PBS→centrifuge→resupend with PBS<br>IJ　Michimori</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td></td></tr>
 +
<tr><td>2</td><td>10/15 colony PCR colony1</td><td></td></tr>
 +
<tr><td>3</td><td>10/15 colony PCR colony2</td><td></td></tr>
 +
<tr><td>4</td><td>10/15 colony PCR colony3</td><td></td></tr>
 +
<tr><td>5</td><td>10/15 colony PCR colony</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/e/ef/T--Kyoto--10_16_No8_Electrophoresis_Sassa_10_17_12_16_22.1.jpg"><img src="https://static.igem.org/mediawiki/2016/a/a2/T--Kyoto--10_16_No8_Electrophoresis_Sassa_10_17_12_16_22.2.jpg
 +
">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Sassa</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>2/22  scFv col.3 0.1</td><td>VF 0.75</td><td>VR 0.75</td><td>KAPA HoFi 12.5</td><td>10.9</td><td></td><td>25</td></tr>
 +
<tr><td>2/22  scFv col.3 0.1</td><td>VF 0.75</td><td>VR 0.75</td><td>KAPA HoFi 12.5</td><td>10.9</td><td></td><td></td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">95℃ 5min<br>98℃ 20s×35<br>55℃ 15s ×35<br>72℃ 30s×35<br>4℃ infinity</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Uchino, Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>10/15 Gel Extraction INP His coding 長</td><td>9.6</td><td>1.88</td><td>0.27</td></tr>
 +
<tr><td>10/15 Gel Extraction INP His coding 短</td><td>74.4</td><td>1.87</td><td>0.84</td></tr>
 +
<tr><td>10/15 Gel Extraction INP His CBDcex</td><td>22.0</td><td>1.97</td><td>0.33</td></tr>
 +
<tr><td>Insert MK</td><td>64.5</td><td>1.92</td><td>1.17</td></tr>
 +
<tr><td>Vector MK</td><td>56.6</td><td>2.37</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>Ligation</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Insert</th><th>Vector</th><th>MilliQ</th><th>Ligation High</th></tr>
 +
<tr><td>10/15 Insert MK/2&microL</td><td>10/15 Gel Extract INP His CBDcex 5.5&microl</td><td>0</td><td>7.5</td></tr>
 +
<tr><td>10/15 Insert MK/1&microL</td><td>10/15 Gel Extract INP His CBDcex 6&microL</td><td>0</td><td>7.5</td></tr>
 +
<tr><td>10/15 Insert MK/1&microL</td><td>10/15 Gel Extract INP His CBDcex 0.5&microL</td><td>6</td><td>7.5</td></tr>
 +
<tr><td>10/15 Digestion INP His coding/5&microL</td><td>10/15 Digestion INP His CBDcex 2.5&microL</td><td>0</td><td>7.5</td></tr>
 +
<tr><td>10/15 Digestion INP His coding/6&microL</td><td>10/15 Digestion INP His CBDcex 1.5&microL</td><td>0</td><td>7.5</td></tr>
 +
<tr><td>10/15 Digestion INP His coding/4.5&microL</td><td>10/15 Digestion INP His CBDcex 3&microL</td><td>0</td><td>7.5</td></tr>
 +
<tr><td>10/14Gel Extract INP His coding/5.5</td><td>10/15 Digestion INP His CBDcex 2&microL</td><td>0</td><td>7.5</td></tr>
 +
<tr><td>10/14Gel Extract INP His coding/6</td><td>10/15 Digestion INP His CBDcex 1.5&microL</td><td>0</td><td>7.5</td></tr>
 +
<tr><td>10/15 Digestion INP His coding 3.5&microL</td><td>10/15 Digestion INP His CBDcex 4&microL</td><td>0</td><td>7.5</td></tr>
 +
<tr><td>10/15 Digestion INP His coding 2.5&microL</td><td>10/15 Digestion INP His CBDcex 5&microL</td><td>0</td><td>7.5</td></tr>
 +
<tr><td>10/15 Digestion INP His coding 3.75</td><td>10/15 Digestion INP His CBDcex 3.75</td><td>0</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>Transformation</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name</th><th>Sample Volume (&microl)</th><th>Competent Cells/(&microl)</th><th>Medium</th></tr>
 +
<tr><td>10/15Ligation Insert MK＋Gel Extract1 2&microL</td><td>2</td><td>DH5α/2</td><td></td></tr>
 +
<tr><td>10/15Ligation Insert MK＋Gel Extract2 2&microL</td><td>2</td><td>DH5α/2</td><td></td></tr>
 +
<tr><td>10/15Ligation Digestion＋Digestion3 2&microL</td><td>2</td><td>DH5α/2</td><td></td></tr>
 +
<tr><td>10/15Ligation Digestion＋Digestion4 2&microL</td><td>2</td><td>DH5α/2</td><td></td></tr>
 +
<tr><td>10/15Ligation Digestion＋Digestion5 2&microL</td><td>2</td><td>DH5α/2</td><td></td></tr>
 +
<tr><td>10/15Ligation Digestion＋Digestion6 2&microL</td><td>2</td><td>DH5α/2</td><td></td></tr>
 +
<tr><td>10/15Ligation Gel Extract＋Digestion7 2&microL</td><td>2</td><td>DH5α/2</td><td></td></tr>
 +
<tr><td>10/15Ligation Gel Extract＋Digestion8 2&microL</td><td>2</td><td>DH5α/2</td><td></td></tr>
 +
<tr><td>10/15Ligation Digestion＋Digestion9 2&microL</td><td>2</td><td>DH5α/2</td><td></td></tr>
 +
<tr><td>10/15Ligation Digestion＋Digestion10 2&microL</td><td>2</td><td>DH5α/2</td><td></td></tr>
 +
<tr><td>10/15Ligation Digestion＋Digestion11 2&microL</td><td>2</td><td>DH5α/2</td><td></td></tr>
 +
<tr><td>10/14 Ligation PuroductA 2&microL</td><td>2</td><td>DH5α/2</td><td></td></tr>
 +
<tr><td>10/14 Ligation PuroductB 2&microL</td><td>2</td><td></td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>PCR (Purification)</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Sample Name/(&microl)</th><th>Concentration(ng/&microl)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>10/15 PCR scFv INP His codingFw＋INP His RV</td><td>284.4</td><td>1.88</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>10/14 Tranceformation INPHis coding2 colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>10/14 Tranceformation INPHis coding2 colony2</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>non-woven felt </h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>sample</th><th>null</th></tr>
 +
<tr><td>cex</td><td>13431×200</td></tr>
 +
<tr><td></td><td>32~41×600</td></tr>
 +
<tr><td>ctl</td><td>13431×200</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>10/14 Trance formation INP His coding 2 colony1</td><td>LB(CP)</td></tr>
 +
<tr><td>10/14 Trance formation INP His coding 2 colony2</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>fluorescence microscope assay</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>sample</th><th>number</th><th>magnification</th></tr>
 +
<tr><td>cex bemliese</td><td>13431</td><td>×200</td></tr>
 +
<tr><td>cex bemliese</td><td>13432~13441</td><td>×600</td></tr>
 +
<tr><td>CTL bemliese</td><td>13442</td><td>×200</td></tr>
 +
<tr><td>CTL bemliese</td><td>13443~13355</td><td>×600</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR</h4></span>
 +
<span class="keln_researcher">Matsumoto, Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>number</th><th>sample</th><th>Any others</th><th>Any others(&microL)</th></tr>
 +
<tr><td>1~5</td><td>10/15Ligation Insert MK＋Gel Extract1</td><td>2×Ready Mix</td><td>12.5</td></tr>
 +
<tr><td>6~10</td><td>10/15Ligation Insert MK＋Gel Extract2</td><td>Fwd primer</td><td>1.25</td></tr>
 +
<tr><td>11~15</td><td>10/15Ligation Insert MK＋Gel Extract3</td><td>Rev primer</td><td>1.25</td></tr>
 +
<tr><td>16~20</td><td>10/15Ligation Digestion＋Digestion4</td><td>MilliQ</td><td>10</td></tr>
 +
<tr><td>21~24</td><td>10/15Ligation Digestion＋Digestion5</td><td></td><td>52×mix</td></tr>
 +
<tr><td>25~28</td><td>10/15Ligation Digestion＋Digestion6</td><td>2×Ready</td><td>650</td></tr>
 +
<tr><td>29~32</td><td>10/15Ligation Gel Extract＋Digestion7</td><td>Fwd </td><td>65</td></tr>
 +
<tr><td>33~36</td><td>10/15Ligation Gel Extract＋Digestion8</td><td>Rev</td><td>65</td></tr>
 +
<tr><td>37~40</td><td>10/15Ligation Digestion＋Digestion9</td><td>MilliQ</td><td></td></tr>
 +
<tr><td>41~44</td><td>10/15Ligation Digestion＋Digestion10</td><td></td><td></td></tr>
 +
<tr><td>45~48</td><td>10/15Ligation Digestion＋Digestion11</td><td></td><td></td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">                                                                      ×16<br>16Colony Picking　DMSO 0.75 12.75 12<br>                                    Mix      12.5 212.5 200<br>                                    Fwd 1.25 21.25 18.75 20<br>                                    Rev 1.25 21.25 18.75 20<br>                                    MilliQ 9.25 157.25 138.75 148<br>16sample colony1~14・・・P15のsample colony<br>Blue　　　　　　　15・・・2/8 INP-His9<br>                                    16・・・8/1 miniprep INPcex colony</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td></td></tr>
 +
<tr><td>2</td><td>1</td><td></td></tr>
 +
<tr><td>3</td><td>2</td><td></td></tr>
 +
<tr><td>4</td><td>3</td><td></td></tr>
 +
<tr><td>5</td><td>4</td><td></td></tr>
 +
<tr><td>6</td><td>5</td><td></td></tr>
 +
<tr><td>7</td><td>6</td><td></td></tr>
 +
<tr><td>8</td><td>7</td><td></td></tr>
 +
<tr><td>9</td><td>8</td><td></td></tr>
 +
<tr><td>10</td><td>9</td><td></td></tr>
 +
<tr><td>11</td><td></td><td></td></tr>
 +
<tr><td>12</td><td></td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/4/4d/T--Kyoto--10_16_No21_Electrophoresis_Sassa_10_17_10_54_10.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td></td><td></td></tr>
 +
<tr><td>2</td><td>1kb ladder</td><td></td></tr>
 +
<tr><td>3</td><td>11</td><td></td></tr>
 +
<tr><td>4</td><td>12</td><td></td></tr>
 +
<tr><td>5</td><td>13</td><td></td></tr>
 +
<tr><td>6</td><td>14</td><td></td></tr>
 +
<tr><td>7</td><td>15</td><td></td></tr>
 +
<tr><td>8</td><td>16</td><td></td></tr>
 +
<tr><td>9</td><td>17</td><td></td></tr>
 +
<tr><td>10</td><td>18</td><td></td></tr>
 +
<tr><td>11</td><td>19</td><td></td></tr>
 +
<tr><td>12</td><td>20</td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/7/76/T--Kyoto--10_16_No21_Electrophoresis_Sassa_10_17_10_54_18.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>Fluorescence Microscopy</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>E.coli</th><th>OD600</th><th>cellulose</th><th>size / weight</th><th>filter</th></tr>
 +
<tr><td>-</td><td>-</td><td>bemliese</td><td>5mm×5mm</td><td>-</td></tr>
 +
<tr><td>INPNC-His-ctl</td><td>1</td><td>bemliese</td><td>5mm×5mm</td><td>-</td></tr>
 +
<tr><td>INPNC-His-CBDcex</td><td>1</td><td>bemliese</td><td>5mm×5mm</td><td>-</td></tr>
 +
<tr><td>-</td><td>-</td><td>powder</td><td>1</td><td>filtered</td></tr>
 +
<tr><td>INPNC-His-ctl</td><td>1</td><td>powder</td><td>1</td><td>filtered</td></tr>
 +
<tr><td>INPNC-His-CBDcex</td><td>1</td><td>powder</td><td>1</td><td>filtered</td></tr>
 +
<tr><td>-</td><td>-</td><td>powder</td><td>0.5</td><td>filtered</td></tr>
 +
<tr><td>INPNC-His-ctl</td><td>1</td><td>powder</td><td>0.5</td><td>filtered</td></tr>
 +
<tr><td>INPNC-His-CBDcex</td><td>1</td><td>powder</td><td>0.5</td><td>filtered</td></tr>
 +
<tr><td>-</td><td>-</td><td>powder</td><td>0.1</td><td>filtered</td></tr>
 +
<tr><td>INPNC-His-ctl</td><td>1</td><td>powder</td><td>0.1</td><td>filtered</td></tr>
 +
<tr><td>INPNC-His-CBDcex</td><td>1</td><td>powder</td><td>0.1</td><td>filtered</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1016" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/16</h3></span>
 +
<span class="keln_exp"><h4>fluorescence microscope assay</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>sample</th><th>number</th><th>magnification</th></tr>
 +
<tr><td>cex 1mg</td><td>13456</td><td>×200</td></tr>
 +
<tr><td>cex 1mg</td><td>13457~13466</td><td>×600</td></tr>
 +
<tr><td>cex 0.5mg</td><td>13467</td><td>×200</td></tr>
 +
<tr><td>cex 0.5mg</td><td>13468~13378</td><td>×600</td></tr>
 +
<tr><td>cex 0.5mg</td><td>13379</td><td>×1000</td></tr>
 +
<tr><td>CTL 1mg</td><td>13380</td><td>×200</td></tr>
 +
<tr><td>CTL 1mg</td><td>13381~13390</td><td>×600</td></tr>
 +
<tr><td>CTL 0.5mg</td><td>13391</td><td>×200</td></tr>
 +
<tr><td>CTL 0.5mg</td><td>13392~13401</td><td>×600</td></tr>
 +
<tr><td>CTL 0.5mg</td><td>13402</td><td>×1000</td></tr>
 +
<tr><td>cex 0.1mg</td><td>13403</td><td>×200</td></tr>
 +
<tr><td>cex 0.1mg</td><td>13404,13407~13416</td><td>×600</td></tr>
 +
<tr><td>cex 0.1mg</td><td>13405,13406</td><td>×1000</td></tr>
 +
<tr><td>CTL 0.1mg</td><td>13417</td><td>×200</td></tr>
 +
<tr><td>CTL 0.1mg</td><td>13418~13430</td><td>×600</td></tr>
 +
<tr><td>CTL 0.1mg</td><td>13431</td><td>×200</td></tr>
 +
<tr><td>cellulose 0.5mg</td><td>13432~13441</td><td>×600</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1017" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/17</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>21</td><td></td></tr>
 +
<tr><td>2</td><td>22</td><td></td></tr>
 +
<tr><td>3</td><td>1kb ladder</td><td></td></tr>
 +
<tr><td>4</td><td>23</td><td></td></tr>
 +
<tr><td>5</td><td>24</td><td></td></tr>
 +
<tr><td>6</td><td>25</td><td></td></tr>
 +
<tr><td>7</td><td>26</td><td></td></tr>
 +
<tr><td>8</td><td>27</td><td></td></tr>
 +
<tr><td>9</td><td>28</td><td></td></tr>
 +
<tr><td>10</td><td>29</td><td></td></tr>
 +
<tr><td>11</td><td></td><td></td></tr>
 +
<tr><td>12</td><td></td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/4/48/T--Kyoto--10_17_No22_Electrophoresis_Sassa_10_17_07_25_05.1.jpg"><img src="https://static.igem.org/mediawiki/2016/9/96/T--Kyoto--10_17_No22_Electrophoresis_Sassa_10_17_07_25_05.2.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1017" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/17</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td></td></tr>
 +
<tr><td>2</td><td>31</td><td></td></tr>
 +
<tr><td>3</td><td>32</td><td></td></tr>
 +
<tr><td>4</td><td>33</td><td></td></tr>
 +
<tr><td>5</td><td>34</td><td></td></tr>
 +
<tr><td>6</td><td></td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/1/13/T--Kyoto--10_17_No23_Electrophoresis_Sassa_10_17_08_53_22.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1017" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/17</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher"></span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>36</td><td></td></tr>
 +
<tr><td>2</td><td>37</td><td></td></tr>
 +
<tr><td>3</td><td>38</td><td></td></tr>
 +
<tr><td>4</td><td>39</td><td></td></tr>
 +
<tr><td>5</td><td></td><td></td></tr>
 +
<tr><td>6</td><td></td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/b/ba/T--Kyoto--10_17_No23_Electrophoresis_Sassa_10_17_08_54_39.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1017" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/17</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td>1kb ladder</td><td></td></tr>
 +
<tr><td>2</td><td>41</td><td></td></tr>
 +
<tr><td>3</td><td>42</td><td></td></tr>
 +
<tr><td>4</td><td>43</td><td></td></tr>
 +
<tr><td>5</td><td></td><td></td></tr>
 +
<tr><td>6</td><td></td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/8/85/T--Kyoto--10_17_No24_Electrophoresis_Sassa_10_17_08_57_02.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1017" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/17</h3></span>
 +
<span class="keln_exp"><h4>Electrophoresis</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Lane</th><th>sample/(&microl)</th><th>Length(bp)</th></tr>
 +
<tr><td>1</td><td></td><td></td></tr>
 +
<tr><td>2</td><td>1kb ladder</td><td></td></tr>
 +
<tr><td>3</td><td>45</td><td></td></tr>
 +
<tr><td>4</td><td>46</td><td></td></tr>
 +
<tr><td>5</td><td>47</td><td></td></tr>
 +
<tr><td>6</td><td></td><td></td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/2/2b/T--Kyoto--10_17_No24_Electrophoresis_Sassa_10_17_08_57_47.jpg
 +
https://static.igem.org/mediawiki/2016/e/ea/T--Kyoto--10_17_No26_Colony_PCR_%28typeA%29_Sassa_10_17_09_47_35.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1017" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/17</h3></span>
 +
<span class="keln_exp"><h4>Liquid Culture</h4></span>
 +
<span class="keln_researcher">Yamamoto</span>
 +
<table class="keln_table">
 +
<tr><th>Name</th><th>medium</th></tr>
 +
<tr><td>colony 1,2,3,4,5,11,12,13,14,15,18,19,22,24,26,30</td><td></td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">total 16mL</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1017" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/17</h3></span>
 +
<span class="keln_exp"><h4>Miniprep</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>DNA</th><th>Concentration/(&microg/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>1</td><td>65.8</td><td>1.91</td><td></td></tr>
 +
<tr><td>2</td><td>63.2</td><td>1.90</td><td>1.84</td></tr>
 +
<tr><td>3</td><td>84.6</td><td>1.87</td><td>1.33</td></tr>
 +
<tr><td>4</td><td>100.8</td><td>1.92</td><td>1.67</td></tr>
 +
<tr><td>5</td><td>84.6</td><td>1.89</td><td>1.59</td></tr>
 +
<tr><td>11</td><td>89.3</td><td>1.84</td><td>1.43</td></tr>
 +
<tr><td>12</td><td>87.3</td><td>1.92</td><td>1.88</td></tr>
 +
<tr><td>13</td><td>88.0</td><td>1.89</td><td>1.59</td></tr>
 +
<tr><td>14</td><td>79.8</td><td>1.91</td><td>1.72</td></tr>
 +
<tr><td>15</td><td>83.3</td><td>1.90</td><td>1.82</td></tr>
 +
<tr><td>18</td><td>102.5</td><td>1.88</td><td>1.50</td></tr>
 +
<tr><td>19</td><td>114.9</td><td>1.91</td><td>1.59</td></tr>
 +
<tr><td>22</td><td>109.9</td><td>1.78</td><td>1.14</td></tr>
 +
<tr><td>24</td><td>80.4</td><td>1.93</td><td>2.00</td></tr>
 +
<tr><td>26</td><td>64.6</td><td>1.93</td><td>1.84</td></tr>
 +
<tr><td>30</td><td>88.8</td><td>1.80</td><td>1.07</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">total 16mL</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1017" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/17</h3></span>
 +
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
 +
<span class="keln_researcher">Yamada</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primers/(&microl)</th><th>MilliQ/(&microl)</th><th>PreMix/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>10/15gel plated colony 1</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>10/15gel plated colony</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>10/15gel plated colony</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>10/15gel plated colony</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>10/15gel plated colony</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>10/15gel plated colony</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>10/15gel plated colony</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>10/15gel plated colony</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>10/15gel plated colony</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>10/15gel plated colony</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>10/15gel plated colony</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>10/15gel plated colony</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>10/15gel plated colony</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>10/15gel plated colony</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>INPNC-His-scFv</td><td>VF1.25 VR1.25</td><td>9.25</td><td>KapaTag/12.5 DMSO 0.75</td><td>25</td></tr>
 +
<tr><td>INP-His-CBDcex</td><td>VF1.25 VR1.25</td><td>9.25</td><td></td><td>25</td></tr>
 +
</table>
 +
</div>
 +
<p class="keln_note">VF 1.25×8=10&microL<br>VR 1.25×8=10&microL<br>Kapa 12.5×8=100&microL<br>DM80 0.75×8=6&microL<br>MilliQ 9.25×8=74&microL<br>Total 200&microL</p>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Sassa -->
 +
<div class="keln_container">
 +
<a name="1017" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/17</h3></span>
 +
<span class="keln_exp"><h4>CBD Binding Assay</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Dual 1mL OD=1.0</th><th>bemlise 5mm×5mm</th><th>VLP Narita104 10&microL</th></tr>
 +
<tr><td>Dual 1mL OD=1.0</td><td>bemlise 5mm×5mm</td><td></td></tr>
 +
<tr><td>ctll 1mL OD=1.0</td><td>bemlise 5mm×5mm</td><td></td></tr>
 +
<tr><td>ctl 1mL OD=1.0</td><td>bemlise 5mm×5mm</td><td></td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1001" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/1</h3></span>
 +
<span class="keln_exp"><h4>RNA purification</h4></span>
 +
<span class="keln_researcher">Ohara</span>
 +
<table class="keln_table">
 +
<tr><th>BclAcex</th><th>0.504</th></tr>
 +
<tr><td>BclAscFv</td><td>0.788</td></tr>
 +
<tr><td>INPscFv</td><td>0.837</td></tr>
 +
<tr><td>INPclos</td><td>1.564</td></tr>
 +
<tr><td>BclAclos</td><td>0.817</td></tr>
 +
<tr><td>INPcex</td><td>0.839</td></tr>
 +
</table>
 +
<img src="https://static.igem.org/mediawiki/2016/d/d1/T--Kyoto--10_1_No161_General_Miyazaki_10_11_10_14_22.jpg">
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1001" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/1</h3></span>
 +
<span class="keln_exp"><h4>RNA extraction</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>number</th><th>sample</th><th>conc(ng/&microl)</th><th>260/280</th><th>260/230</th><th>diluted solution volume(200ng/&microl)</th></tr>
 +
<tr><td>1</td><td>INPclos</td><td>359.2</td><td>2.07</td><td>1.21</td><td>11.94</td></tr>
 +
<tr><td>2</td><td>BclAcex</td><td>230.9</td><td>2.03</td><td>0.85</td><td>2.31</td></tr>
 +
<tr><td>3</td><td>INPcex</td><td>1069.0</td><td>2.11</td><td>1.43</td><td>65.17</td></tr>
 +
<tr><td>4</td><td>INPscFv</td><td>320.0</td><td>2.10</td><td>0.93</td><td>9</td></tr>
 +
<tr><td>5</td><td>BclAclos</td><td>386.2</td><td>2.04</td><td>0.97</td><td>13.97</td></tr>
 +
<tr><td>6</td><td>BclAcex</td><td>441.0</td><td>2.04</td><td>0.83</td><td>18</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1001" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/1</h3></span>
 +
<span class="keln_exp"><h4>Reverse Transcription</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>name</th><th>copy(high or low)</th><th>RT</th></tr>
 +
<tr><td>3 nega</td><td>high copy</td><td>-</td></tr>
 +
<tr><td>3 posi</td><td>high copy</td><td>+</td></tr>
 +
<tr><td>4 nega</td><td>low copy</td><td>-</td></tr>
 +
<tr><td>4 posi</td><td>low copy</td><td>+</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1001" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/1</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>10/1 RT×20 3RT- /1.0&microl</td><td>rRNA16s forward primer1 /0.4&microl</td><td>rRNA16s reverse primer1 /0.4&microl</td><td>2×KAPA HiFi /5&microl</td><td>3.2&microl</td><td></td><td>10&microl</td></tr>
 +
<tr><td>10/1 RT×20 3RT+ /1.0&microl</td><td>rRNA16s forward primer1 /0.4&microl</td><td>rRNA16s reverse primer1 /0.4&microl</td><td>2×KAPA HiFi /5&microl</td><td>3.2&microl</td><td></td><td>10&microl</td></tr>
 +
<tr><td>10/1 RT×20 4RT- /1.0&microl</td><td>rRNA16s forward primer1 /0.4&microl</td><td>rRNA16s reverse primer1 /0.4&microl</td><td>2×KAPA HiFi /5&microl</td><td>3.2&microl</td><td></td><td>10&microl</td></tr>
 +
<tr><td>10/1 RT×20 4RT+ /1.0&microl</td><td>rRNA16s forward primer1 /0.4&microl</td><td>rRNA16s reverse primer1 /0.4&microl</td><td>2×KAPA HiFi /5&microl</td><td>3.2&microl</td><td></td><td>10&microl</td></tr>
 +
<tr><td>9/27 RT×1 A-6 /1.0&microl</td><td>rRNA16s forward primer1 /0.4&microl</td><td>rRNA16s reverse primer1 /0.4&microl</td><td>2×KAPA HiFi /5&microl</td><td>3.2&microl</td><td></td><td>10&microl</td></tr>
 +
<tr><td>9/27 RT×1 A-5 /1.0&microl</td><td>rRNA16s forward primer1 /0.4&microl</td><td>rRNA16s reverse primer1 /0.4&microl</td><td>2×KAPA HiFi /5&microl</td><td>3.2&microl</td><td></td><td>10&microl</td></tr>
 +
<tr><td>9/27 RT×1 B-6 /1.0&microl</td><td>rRNA16s forward primer1 /0.4&microl</td><td>rRNA16s reverse primer1 /0.4&microl</td><td>2×KAPA HiFi /5&microl</td><td>3.2&microl</td><td></td><td>10&microl</td></tr>
 +
<tr><td>9/27 RT×1 B-5 /1.0&microl</td><td>rRNA16s forward primer1 /0.4&microl</td><td>rRNA16s reverse primer1 /0.4&microl</td><td>2×KAPA HiFi /5&microl</td><td>3.2&microl</td><td></td><td>10&microl</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: Miyazaki -->
 +
<div class="keln_container">
 +
<a name="1001" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/1</h3></span>
 +
<span class="keln_exp"><h4>PCR (Target)</h4></span>
 +
<span class="keln_researcher">Uchino</span>
 +
<table class="keln_table">
 +
<tr><th>Templates/(&microl)</th><th>Primer1/(&microl)</th><th>Primer2/(&microl)</th><th>Buffer/(&microl)</th><th>MilliQ/(&microl)</th><th>Any Other/(&microl)</th><th>Total/(&microl)</th></tr>
 +
<tr><td>10/1 RT×20 3RT- /1.0</td><td>INP primer for CBD /0.4</td><td>CBD cex primer Rv /0.4</td><td>2×KAPA HiFi /5</td><td>3.2</td><td></td><td>10</td></tr>
 +
<tr><td>10/1 RT×20 3RT+ /1.0</td><td>INP primer for CBD /0.4</td><td>CBD cex primer Rv /0.4</td><td>2×KAPA HiFi /5</td><td>3.2</td><td></td><td>10</td></tr>
 +
<tr><td>10/1 RT×20 4RT- /1.0</td><td>scFv primer1 /0.4</td><td>scFv primer2 /0.4</td><td>2×KAPA HiFi /5</td><td>3.2</td><td></td><td>10</td></tr>
 +
<tr><td>10/1 RT×20 4RT+ /1.0</td><td>scFv primer1 /0.4</td><td>scFv primer2 /0.4</td><td>2×KAPA HiFi /5</td><td>3.2</td><td></td><td>10</td></tr>
 +
<tr><td>9/27 RT×1 A-6 /1.0</td><td>INP primer for CBD /0.4</td><td>CBD cex primer Rv /0.4</td><td>2×KAPA HiFi /5</td><td>3.2</td><td></td><td>10</td></tr>
 +
<tr><td>9/27 RT×1 A-5 /1.0</td><td>INP primer for CBD /0.4</td><td>CBD cex primer Rv /0.4</td><td>2×KAPA HiFi /5</td><td>3.2</td><td></td><td>10</td></tr>
 +
<tr><td>9/27 RT×1 B-6 /1.0</td><td>scFv primer1 /0.4</td><td>scFv primer2 /0.4</td><td>2×KAPA HiFi /5</td><td>3.2</td><td></td><td>10</td></tr>
 +
<tr><td>9/27 RT×1 B-5 /1.0</td><td>scFv primer1 /0.4</td><td>scFv primer2 /0.4</td><td>2×KAPA HiFi /5</td><td>3.2</td><td></td><td>10</td></tr>
 +
</table>
 +
</div>
 +
<!------------ Table END ------------>
  
−
<span class="keln_researcher">Sukegawa</span>
 
  
−
<table class="keln_table">
 
  
−
<tr><th>Templates/(μl)</th><th>Primer1/(μl)</th><th>Primer2/(μl)</th><th>Buffer/(μl)</th><th>MilliQ/(μl)</th><th>Any Other/(μl)</th><th>Total/(μl)</th></tr>
 
  
−
<tr><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
 
  
−
</table>
 
  
−
</div>
 
  
−
<!------------ Table END ------------>
 
  
 +
<!-- CONTENTS_HERE -->
 
     </div>
 
     </div>
 
     </div>
 
     </div>

Revision as of 11:17, 18 October 2016

8/21

Liquid Culture

 Li
Namemedium
strainMedium/ml
8/20 transformation RBSPG(AMP) /6
8/20 dilution INP7PG(CP) /6
8/17 transformation 3c5, RFPPG(CP) /6
8/20 transformation p+ [1]PG(AMP) /6
8/20 transformation p- [1]PG(AMP) /6

8/21

Electrophoresis

 Li
Lanesample/(&microl)Length(bp)
11kb ladder /1.5
28/21 cloning PCR scfv[1] /2
38/21 cloning PCR scfv[2] /2
4nega /2
58/21 colony PCR p+ [1] /2
68/21 colony PCR p+ [2] /2
78/21 colony PCR p+ [3] /2
88/21 colony PCR p- [1] /2
98/21 colony PCR p- [2] /2
108/21 colony PCR p- [3] /2
118/21 colony PCR parts p+ /2
128/21 colony PCR parts p- /2

[img35]
2, 3, 4 cloning PCR failed.
1 forgot to put ladder
5, 7, 8, 9 strong band
6 weak
10 failed
P+[1] and p-[1] -> liquid culture

8/24

Ligation

 Li
sample/&microlMilliQ (&microl)Buffer /&microlLigase(&microl)Total(&microl)
8/19 mutation PCR scfv /53.510.510

8/24

Liquid Culture

 Li
Namemedium
8/20 transformation m.scfv colonyA /6PG(AMP)

8/24

Miniprep

 Li
DNAConcentration/(&microg/ml)260/280260/230
8/20 Liquid culture RBS124.51.651.02
8/20 Liquid culture INP147.81.660.98
8/20 Liquid culture 3c5.RFP234.81.791.62
8/20 Liquid culture P+[1]40.31.781.60
8/20 Liquid culture P-[1]56.71.802.05

8/25

Miniprep

 Li, Uchino, Notsu, Kaneko
DNAConcentration/(&microg/ml)260/280260/230
8/24 LIquid Culture m.scfv colonyA---
8/24 LIquid Culture m.scfv colonyB--

8/25

Electrophoresis
Lanesample/(&microl)Length(bp)
11kb ladder /1.5
28/24 miniprep p+(J61002) /2
38/24 miniprep p-(J61002) /2
48/24 miniprep RFP (3c5) /2
58/24 miniprep INP (1c3) /2
68/24 miniprep RBS (1A2) /2

[img38]

8/25

Restriction Enzyme Digestion

 Uchino
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
p+/24.81110xH /49.240
IINP /6.81110xM /427.240
p- /17.61110xH /416.440
RBS /8.01110xH /42640
RFP /4.31110xH /429.740

8/25

Gel Extraction

 Uchino, Notsu
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder 6
2, 38/25 digestion p+24 each
4, 58/25 digestion p- 24 each
6, 78/25 digestion RFP 24 each
8, 98/25 digestion INP 24 each
10, 118/25 digestion RBS 24 each

[img40]

8/25

Actvation check of Xba and Pst

 Uchino
INP(&microl)Xba(&microl)Pst(&microl)10xM(&microl)BSA(&microl)MilliQ(&microl)Total(&microl)
0.680.10.110.27.9210
0.680.1-10.28.0210
0.68-0.110.28.0210
0.68--10.28.1210

8/25

Electrophoresis

 Notsu
Lanesample/(&microl)Length(bp)
11kb ladder /3
2[1] (inp xp cut) /10
3[2] (inp x cut) /10
4[3] (inp p cut) /10
5[4] (inp no cut) /10

[img42]

8/26

Restriction Enzyme Digestion

 Uchino, Notsu
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
8/24 miniprep INP 1c3 /0.68-1.00.27.1210
8/15 miniprep RFP psb1c3 /1.3-1.00.26.510
8/24 miniprep INP 1c3 /0.680.11.00.27.0210
8/15 miniprep RFP psb1c3 /1.30.11.00.26.410
8/24 miniprep INP 1c3/ 0.680.1-0.28.0210
8/15 miniprep RFP psb1c3 /1.30.1-0.27.410
8/24 miniprep INP 1c3 /0.68--0.28.1210
8/15 miniprep RFP psb1c3 /1.3--0.27.510

8/26

PCR (Target)

 Li
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
scfv from MKc.scfv fw/0.75c.scfv rv/0.752x KAPA Hifi /12.51125
nonec.scfv fw/0.75c.scfv rv/0.752x KAPA Hifi /54.410

8/26

Activation Check of Xba

 Uchino
Lanesample/(&microl)Length(bp)
11kb ladder/2
2[8] RFP/10
3[6] RFP EcoRI/10
4[2] RFP XbaI/10
5[4] RFP EcoRI, XbaI /10
6[7] INP /10
7[5] INP EcoRI /10
8[1] INP XbaI /10
9[3] INP EcoRI, XbaI/10

[img45-1]
[img45-2]

8/27

Electrophoresis

 Li
Lanesample/(&microl)Length(bp)
21kb ladder2
38/25 PCR scfv5
48/25 PCR none5

8/27

PCR (Steps)

 Li
StepsTemparatureTimeCycle
Predenature953min1
Denature9820sec25
Annealing6515sec25
Extension7230sec25
Final Elogation725min1
Final Holding4-

8/27

PCR (Target)

 Li
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
s.BANBAN.Fw/ 0.75BAN.Rv/ 0.75112xKAPA HiFi/ 12.525
s.BANBAN.Fw/ 0.75BAN.Rv/ 0.75112xKAPA HiFi/ 12.525
s.BANBAN.Fw/ 0.75BAN.Histag/ 0.75112xKAPA HiFi/ 12.525
scfvscfv.Fw/ 0.75scfv.Rv/ 0.75112xKAPA HiFi/ 12.525
8/24 miniprep INPINP.Fw/ 0.75INP.Rv/ 0.75112xKAPA HiFi/ 12.525
8/24 miniprep INPINP.Fw/ 0.75INP.Rv.Histag/ 0.75112xKAPA HiFi/ 12.5

8/14

Transformation

 Li, Uchino, Kaneko
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
RFP 0.5ng/&microl2DH5α /20LB(CP)
RFP 5ng/&microl2DH5α /20LB(CP)
RFP 10ng/&microl2DH5α /20LB(CP)
RFP 20ng/&microl2DH5α /20LB(CP)
RFP 50ng/&microl2DH5α /20

8/14

Gel Extraction

 Li, Uchino, Kaneko
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
110/1 PQ(3)about 23.5&microl
21kb ladder6
310/1 PQ(3)about 23.5&microl
410/1 PQ(3)about 24&microl

Total volume of samples of lane 1~3 was 71.5&microl and it was divided into three.

8/14

Transformation

 Li, Uchino, Kaneko
Sample NameSample VolumeCompetent Cells/(&microl)Medium
3C510&microlDH5留 20&microlLB(CP)
P+10&microlDH5留 20&microlLB(CP)
P-10&microlDH5留 20&microlLB(CP)
RBS10&microlDH5留 20&microl

8/14

Liquid Culture

 Li, Uchino, Iguchi, Kaneko
Namemedium
8/13 transformation RFP 20ng/&microl colony1LB(CP)
8/13 transformation part RFP colony1LB(CP)
8/13 transformation part RFP colony2LB(CP)
8/13 transformation part RFP colony3LB(CP)
8/13 transformation part RFP colony4LB(CP)
8/13 transformation part RFP colony5LB(CP)
8/13 transformation part RFP colony6LB(CP)
8/13 transformation part RFP colony7LB(CP)
8/13 transformation part RFP colony8LB(CP)
8/13 transformation part RFP colony9LB(CP)
8/13 transformation part RFP colony10LB(CP)
LB(CP)

8/15

Miniprep

 Li, Uchino, Notsu, Kaneko, Watanabe
DNAConcentration/(&microg/ml)260/280260/230
RFP1 /2&microl82.51.401.22
RFP2 /4&microl142.51.692.05
RFP3 /4&microl501.551.71
RFP4 /4&microl57.51.641.88
RFP6 /4&microl98.751.711.40
RFP(20) /4&microl76.251.67

8/15

Electrophoresis

 Notsu, Kaneko
LaneRestriction Enzyme Digestion Products
11kb ladder 3&microl
2RFP1 10&microl
3RFP2 10&microl
4RFP3 10&microl
5RFP4 10&microl
6RFP6 10&microl
7RFP(20) 10&microl
8PCR1 5&microl
9PCR2 5&microl
10PCR3 5&microl
11PCR4 5&microl
12 PCR5 5 &microl
131kb ladder 3&microl

8/15

Restriction Enzyme Digestion

 Uchino, Notsu, Kaneko
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
8/15Miniprep PSB1C3-RFP12410015(H)01950
8/15Miniprep PSB1C3-RFP21410015(H)02950
8/15Miniprep PSB1C3-RFP34010015(H)0350
8/15Miniprep PSB1C3-RFP43510015(H)0850
8/15Miniprep PSB1C3-RFP62010015(H)02350

8/15

colony PCR

 Uchino, Notsu, Kaneko, Watanabe
colony Primers
8/14 transformation pSB1C3-RFP1 VF+VR
8/14 transformation pSB1C3-RFP2 VF+VR
8/14 transformation pSB1C3-RFP3 VF+VR
8/14 transformation pSB1C3-RFP4 VF+VR
8/14 transformation pSB1C3-RFP1 VF+VR

8/15

PCR (Steps)

 Uchino, Notsu, Kaneko, Watanabe
StepsTemparatureTimeCycle
Pre Denature94℃2min1
Denature94℃30sec30
Annealing55℃30sec30
Extension68℃1min30
Final elongation4℃7min
Final Holding4℃--

8/16

Gel Extraction

 Li, Iguchi, Notsu, Kaneko, Watanabe
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder10
2blank
38/15 digestion pSB1C3-RFP 155
48/15 digestion pSB1C3-RFP 255
58/15 digestion pSB1C3-RFP 336
68/15 digestion pSB1C3-RFP 436
78/15 digestion pSB1C3-RFP 6

8/16

Electrophoresis

 Li, Iguchi, Notsu, Kaneko, Watanabe
Lanesample/(&microl)Length(bp)
18/16 Gel Extraction RFP1 /1
28/16 Gel Extraction RFP2 /1
38/16 Gel Extraction RFP3 /1
48/16 Gel Extraction RFP4 /1
5blank
68/16 pSB1C3-vector /1
78/16 pSB1C3-vector /1
8

8/16

Ligation

 Li, Iguchi, Notsu, Kaneko, Watanabe
Vector DNA /(&microl) insert DNA /(&microl) ligase /(&microl) 10×buffer /(&microl) MilliQ /(&microl) Total /(&microl)
pSB1C3 EPcut /1 RFP /4 0.5 1 3 9.5
pSB1C3 EPcut /1 RFP /2 0.5 1 5 9.5
pSB1C3 EPcut /1 RFP /4 0.5 1 3 9.5
pSB1C3 EPcut /1 RFP /4 0.5 1 3 9.5
pSB1C3 EPcut /1 - 0.5 1 7 9.5

8/17

Liquid Culture

 Li
Namemedium
8/16 transformation pSB3C5 RFP1LB(CP) /6
8/16 transformation pSB3C5 RFP2LB(CP) /6
8/16 transformation pSB3C5 RFP1LB(CP) /6

8/17

Transformation

 Li
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
pSB1C3 P+2DH5α /20LB
pSB1C3 P-2DH5α /20LB
pSB1C3 RBS2DH5α /20LB
pSB3C5 RFP2DH5α /20LB
8/16 ligation pSB1C3 RFP12DH5αLB

8/17

Electrophoresis

 Iguchi
Lanesample/(&microl)Length(bp)
11kbp ladder /3
2blank
3pSB1C3-RFP(ligate)1 /2
4pSB1C3-RFP(ligate)2 /2
5pSB1C3-RFP(ligate)3 /2
6pSB1C3-RFP(ligate)4 /3
7Vector (Ctrl) /5

8/18

Liquid Culture

 Li, Iguchi, Notsu
Namemedium
INP [1] BBa_K811005LB(CP) / 6ml
INP [2] BBa_K811005LB(CP) / 6ml
INP [3] BBa_K811005LB(CP) / 6ml
scfv04 [1] BBa_K875004LB(CP) / 6ml
scfv04 [2] BBa_K875004LB(CP) / 6ml
scfv04 [3] BBa_K875004LB(CP) / 6ml
scfv06 [1] BBa__K875006LB(CP) / 6ml
scfv06 [2] BBa_K875006LB(CP) / 6ml
scfv06 [3] BBa_K875006LB(CP) / 6ml

8/18

Miniprep

 Li, Iguchi, Notsu
DNAConcentration/(&microg/ml)260/280260/230
8/17 Liquid Culture 3C5 [1] / 5&microl771.691.45
8/17 Liquid Culture 3C5 [2] / 2&microl401.671.17
8/17 Liquid Culture 3C5 [3] / 2&microl153.753.70

8/18

Electrophoresis

 Li
Lanesample/(&microl)Length(bp)
11kb ladder /1-
2[1] psb3c5(8/18 miniprep) /4.23807
3[2] psb3c5(8/18 miniprep) /4.23807
4[1'] psb3c5(8/18 miniprep) /4.23807
5[2'] psb3c5(8/18 miniprep) /4.23807
6[3'] psb3c5(8/18 miniprep) /4.23807
https://static.igem.org/mediawiki/2016/c/c5/T--Kyoto--8_18_No18_Electrophoresis_Kaneko_05_20_04_12_53.jpg

[img18]

8/18

Restriction Enzyme Digestion

 Li, Iguchi, Notsu
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
psb3c5(8/18miniprep) [1'], [2'], [3'] mixed / 501.01.05.656.6

8/19

Transformation

 Li, Iguchi, Notsu
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
BBa_ J61002 p+2DH5a /20LB(Amp)
BBa_J61002 p-2DH5a /20LB(Amp)
BBa_1A2 RBS2DH5a /20LB(Amp)

Failed due to deactivation of Amp

8/19

MutationPCR
Templates /&microlPrimer /&microlPrimer STAR Max Premix /&microlMilliQ /&microlTotal(&microl)
scfv plasmid /0.1m.scfv FW /1, m.scfvRV/12522.950
scfv plasmid (nega) /0.1054.910
(nega) /0m.scfv FW/0.2, m.scfvRV/0.254.610

---30cycle---
98℃ 10''
55℃ 15''
72℃ 20''
----------------
4℃ holding

8/19

Electrophoresis

 Notsu
Lanesample/(&microl)Length(bp)
11kb ladder/2-
2scfv plasmid +primer(m.scfv FW, m.scfv RV) mutation PCR product /53600
3scfv plasmid mutation PCR product /5-
4primer(m.scfv FW, m.scfv RV) mutation PCR product /5-
5psb3c5 EPcut product /52600(backborn), 1200(inseert)

[img22]

8/19

Concentration

 Li, Notsu
SampleOD600Bacterial cells /ml
INP [1] K8110050.6435.14x10^8
scfv06 K8750060.6345.07x10^8
scfv04 K8750040.6184.94x10^8

1 OD 600 equals to 8x10^8 cells /ml

8/19

Miniprep

 Li, Notsu
DNAConcentration/(&microg/ml)260/280260/230
scfv 04[1] K875004 / 3&microl561.631.45
scfv 04[2] K875004 / 3&microl
scfv 04[3] K875004 / 3&microl
scfv 06[1] K875006 /3&microl
scfv 06[2] K875006 /3&microl
scfv 06[3] K875006 /3&microl
INP [1] K811005 /3&microl
INP [2] K811005 /3&microl
INP [3] K811005 /3&microl

There was a problem with an instrument

8/20

Transformation

 Li, Uchino
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
8/19 mutation PCR scfv plasmid (nega)1DH5a/ 20LB(CP)

8/19 mutation PCR:
1&microl extraction of scfv plasmid (nega)/10&microl was folded 300 times.

8/20

Transformation

 Li, Uchino
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
8/19 mutation PCR scfv plasmid1 [note:1]DH5a/20LB(AMP)
8/19 mutation PCR scfv plasmid (nega)1 [note:2]DH5a/20LB(AMP)
BBa_J61002 p+1DH5a/20LB(AMP)
BBa_J61002 p-1DH5a/20LB(AMP)
BBa_1A2 RBS1DH5a/20LB(AMP)

[note:1]
8/19 mutation PCR:
1&microl extraction of scfv plasmid (nega)/50&microl was folded 300 times.
[note:2]
8/19 mutation PCR:
1&microl extraction of scfv plasmid (nega)/10&microl was folded 300 times.

8/20

Electrophoresis

 Li
Lanesample/(&microl)Length(bp)
11kb ladder /1.5
29/19 miniprep scfv04 [1]
39/19 miniprep scfv04 [2]
49/19 miniprep scfv04 [3]
59/19 miniprep scfv06 [1]
69/19 miniprep scfv06 [2]
79/19 miniprep scfv06 [3]
89/19 miniprep INP [1]
99/19 miniprep INP [2]
109/19 miniprep INP [3]

[img26]

8/21

Colony PCR

 Uchino
ColonyPrimers
8/20 transformation RBSVF+VR
8/20 transformation p+VF+VR
8/20 transformation p-VF+VR
8/20 dilution INPVF+VR

8/21

PCR (Steps)

 Uchino
StepsTemparatureTimeCycle
Pre94℃2min1
De94℃30 sec30
An55℃30 sec30
Ex68℃1min30
Final E4℃7min1
Final Hold4℃--

Autoclaved distrilled water /23&microl
2xQuick taq /25&microl
10 pmol/&microl primer (VF) /1&microl
10 pmol/&microl primer (VR) /1&microl
Template DNA /0&microl
Total 50&microl

8/21

Electrophoresis

 Li
Lanesample/(&microl)Length(bp)
11kb ladder /2
28/21 colony PCR RBS /all
38/21 colony PCR p+ /all
48/21 colony PCR p- /all
58/21 colony PCR INP /all
68/21 colony PCR Nega /all

[img29-1]
weak band except INP -> retry
colony PCR and Electrophoresis were conducted under the same condition.
[img29-2]
RBS and INP success.
P+ and P- failed -> retry.

8/21

PCR (Steps)

 Li
StepsTemparatureTimeCycle
Predenature95℃3min1
Denature98℃20sec25
Annealing70℃15sec25
Extension72℃30sec25
Final Elongation72℃5min1
Final Holding4℃-1

8/21

PCR (Target)

 Li
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
scfv 8/20 transformationcscfv.f /0.75cscfv.r /0.752x KAPA Hifi /12.51125
- (nega)cscfv.f /0.3cscfv.f /0.32x KAPA Hifi /54.410

8/21

Colony PCR

 Li
ColonyPrimers
8/20 transformation p+ [1]VF+VR
8/20 transformation p+ [2]VF+VR
8/20 transformation p+ [3]VF+VR
8/20 transformation p- [1]VF+VR
8/20 transformation p- [2]VF+VR
8/20 transformation p- [3]VF+VR
8/18 parts p+VF+VR
8/18 parts p-VF+VR

8/21

PCR (Steps)

 Li
StepsTemparatureTimeCycle
pre94℃2min1
de94℃30sec30
an55℃30sec30
ex68℃1min30
F.elong4℃7min1
F.hold4℃-1

9/8

Restriction Enzyme Digestion

 Notsu
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
[1] 8/15 miniprep 1c3 RFP /140.50.5-10xH /31230
[2] 8/24 miniprep 3c5 RFP /8.50.5-0.510xH /317.530
[3] 8/24 miniprep 3c5 RFP /8.50.50.5-10xH /317.530
[4] 8/24 miniprep BBa_J61002 p- /240.5-0.510xH /3230

9/9

Electrophoresis

 Notsu
LaneRestriction Enzyme Digestion Products
1100bp ladder 9&microL
28/24 miniprep p- EPcut 30&microL

9/9

Electrophoresis

 Sukegawa, Notsu
LaneRestriction Enzyme Digestion Products
11kbp ladder 9&microL
28/15 miniprep 1C3RFP EScut 30&microL
38/24 miniprep 3C5RFP EPcut 30&microL
48/24 miniprep 3C5RFP EScut 30&microL

9/14

PCR (Steps)

 Li
StepsTemparatureTimeCycle
1953min１
29870sec25
36815sec25
47230sec25
5725min1
610∞1

9/14

PCR (Target)

 Li
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
8/24 miniprep INPINPfw/0.75INPRv/0.752×KAPAHiFi1125
8/24 miniprep INPINPfw/0.75INPHisRv/0.752×KAPAHiFi1125
BANBANFw/0/75BANRv/0.752×KAPAHiFi1125
BANBANFw/0/75BANHisRv/0.752×KAPAHiFi1125
BAN BANFw/0/75RBSRv/0.752×KAPAHiFi1125
scFv fromMkscFvFw/0/75scFvRv/0/752×KAPAHiFi1125
scFv fromMkscFvHisFw/0.752×KAPAHiFi1125

9/14

Gel Extraction

 Kaneko
LaneRestriction Enzyme Digestion Products
2ladder/6
49/14 PCR1/10
69/14 PCR2/10
89/14 PCR3/10
109/14 PCR4/10
129/14PCR5/10
149/14 PCR6/10
169/14 PCR7/10

9/14

PCR (Target)

 Li
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3sBAN/0.15+6scFvfromMK/0.15BANFw/0/75scFvRv/0.752×KAPAHiFi/12.525
4sBAN/0.15+7scFvfromMK/0.15BANFw/0/75scFvRv/0.752×KAPAHiFi/12.525
1INP8/24miniprep/0.15+5sBAN0.15+scFvfromMK/0.15BANFw/0/75scFvRv/0.752×KAPAHiFi/12.525
2INP8/24miniprep/0.15BANFw/0/75scFvRv/0.752×KAPAHiFi/12.525

9/15

Restriction Enzyme Digestion

 Li, Uchino
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
3sBAN+6scFvfromMKPCRproduct3411440
4sBAN+7scFvfromMK PCRproduct3411440
1INP8/24miniprep+5sBAN+6scFvfromMKPCRproduct1611220
2INP8/24miniprep+5sBAN+7scFvfromMk PCRproduct11440

9/15

Gel Extraction

 Li, Uchino
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder
23sBAN+6scFvfromMkPCR産物EScut20
33sBAN+6scFvfromMkPCR産物EScut20
44sBAN+7scFvfromMkPCR産物EScut20
54sBAN+7scFvfromMkPCR産物EScut20
61INP8/24miniprep+5sBAN+7scFvfromMkPCR産物EScut20
72INP8/24miniprep+5sBAN+7scFvfromMkPCR産物EScut20
82INP8/24miniprep+5sBAN+7scFvfromMkPCR産物EScut20
9RFP3C5PCR産物

9/16

Colony PCR (2Column)

 Uchino, Michimori
Name IName II
9/15 transformation BANscFv 3C5 colony1~16VF+VR
9/15 transformation INPscFv 3C5 colony1~16VF+VR
9/15 transformation BANHisscFv 3C5 colony1~16VF+VR
9/15 transformation INPHisscFv 3C5 colony1~16

9/16

PCR (Steps)

 Uchino, Michimori
StepsTemparatureTimeCycle
19421
2940.525
3550.525
468125
5471
64∞

9/16

Gel Extraction

 Li, Kaneko, Notsu
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder5
2BANscFv3C5 colony 910
3BANscFv3C5 colony 1010
4BANscFv3C5 colony 1110
5BANscFv3C5 colony 1210
6BANscFv3C5 colony 1310
7BANscFv3C5 colony 1410
8BANscFv3C5 colony 1510
9BANscFv3C5 colony 1610
10BANHisscFv3C5 colony 910
11BANHisscFv3C5 colony 1010
12BANHisscFv3C5 colony 1110
13BANHisscFv3C5 colony 1210
14BANHisscFv3C5 colony 1310
15BANHisscFv3C5 colony 1410
16BANHisscFv3C5 colony 1510
17BANHisscFv3C5 colony 1610
18BANscFv3C5 colony 110

9/16

Gel Extraction

 Li, Kaneko, Notsu
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder5
2INPscFv3C5 colony 910
3INPscFv3C5 colony 1010
4INPscFv3C5 colony 1110
5INPscFv3C5 colony 1210
6INPscFv3C5 colony 1310
7INPscFv3C5 colony 1410
8INPscFv3C5 colony 1510
9INPscFv3C5 colony 1610
10INPHisscFv3C5 colony 910
11INPHisscFv3C5 colony 1010
12INPHisscFv3C5 colony 1110
13INPHisscFv3C5 colony 1210
14INPHisscFv3C5 colony 1310
15INPHisscFv3C5 colony 1410
16INPHisscFv3C5 colony 1510
17INPHisscFv3C5 colony 1610
18BANscFv3C5 colony 110

9/17

Electrophoresis

 Michimori, Kaneko
LaneRestriction Enzyme Digestion Products
1buffer/5
21kb ladder/5
3BAN 9 P/4.5
4BAN 9 N/4.5
5BAN14 P/4.5
6BAN14 N/4.5
7BANHis14 P/4.5
8BANHis14/4.5

9/17

Electrophoresis

 Kaneko
Lanesample/(&microl)Length(bp)
11kbp ladder
2none
3BANHis16 p
4BANHis16 n
5INP 12 p
6INP 14 p
7INP 12 n
8

9/17

Electrophoresis

 Kaneko
LaneRestriction Enzyme Digestion Products
1-
2-
31kbladder/5
4INPHis9 P/4.5
5INPHis9N/4.5
6INPHis12P/4.5
7INPHis12N/4.5
8INP14 N/4.5

9/17

Restriction Enzyme Digestion

 Uchino, Michimori
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
BAN9 20.50.510×H/1610
BAN9 10010×H/1810
BAN14 20.50.510×H/1610
BAN14 10010×H/1810
BANHis14 20.50.510×H/1610
BANHis 14 10010×H/1810
BANHis 16 20.50.510×H/1610
BANHis16 10010×H/1810
INP12 20.50.510×H/1610
INP12 10010×H/1810
INP14 20.50.510×H/1610
INP 14 10010×H/1810
INP 9 20.50.510×H/1610
INP9 10010×H/1810
INPHis12 20.50.510×H/1610
INPHis12 10010×H/1810

9/17

Miniprep

 Notsu
DNAConcentration/(&microg/ml)260/280260/230
9/16 liquid culture BANscFv12
9/16 liquid culture BANscFv14
9/16 liquid culture BANHisscFv12
9/16 liquid culture BANHisscFv14
9/16 liquid culture INPHisscFv12
9/16 liquid culture INPHisscFv14
9/16 liquid culture INPHisscFv10
9/16 liquid culture BANscFv

2/4

PartsAwaking

 Kaneko, Wan
DescriptionPartNameBackboneWellPlateYearPlateNumberMilliQ(&microl)
RFPE1010pSB1C311N20153

2/4

Transformation

 Uchino, Iguchi, Notsu, Wan
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
E10102DH5αLB(CP)

2/5

Colony PCR (typeA)

 Uchino, Egashira, Yamamoto
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
2/4 transformation E1010 colony1VF 6.25&microl VR 6.25&microl12.52×Quick Taq HS DyeMix 25&microl50

2/5

PCR (Steps)

 Uchino, Egashira, Yamamoto
StepsTemparatureTimeCycle
Predenature94℃2min1
Denature94℃30sec25
Annealing51℃30sec25
Extension68℃90sec25
Final Elongation4℃--
Final Holding4℃-

2/5

Transformation

 Uchino
Sample NameSample VolumeCompetent Cells/(&microl)Medium
9/17 miniprep BAN92DH5α/20&microL
9/17　miniprep BANHis142DH5α/20&microL
9/17miniprep INPHis122DH5α/20&microL
9/17miniprep INPHis92

2/5

Liquid Culture

 Uchino, Wan, Yamamoto
Namemedium
2/4 transformation E1010 colony1

2/5

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
11kb ladder /5
22/5 colony PCR E10101019
3other sample-
4other sample-
5other sample-
6other sample-
7other sample-
8other sample

2/6

Miniprep

 Notsu, Kaneko, Egashira, Wan, Yamamoto
DNAConcentration/(&microg/ml)260/280260/230
2/5 liquid culture E1010(RFP) colony148.01.93

2/6

Restriction Enzyme Digestion

 Kaneko, Egashira, Wan, Yamamoto
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
2/6 miniprep E1010 colony13401011×M /40040

2/7

Gel Extraction

 Kaneko, Yamamoto
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder 10
2E1010 colony1 2/6 XPcut40
3other sample-
4other sample-

2/7

Gel Extraction(Measurement)

 Kaneko, Yamamoto
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
E1010 colony1 2/6 XPcut8.21.030.49

2/7

Liquid Culture

 Uchino
Namemedium
2/5 transformation BAN9 colony1LB(CP)
2/5 transformation BAN-HIS14 colony1LB(CP)
2/5 transformation INP-HIS12 colony1LB(CP)
2/5 transformation BAN9 colony1LB(CP)

2/8

Liquid Culture

 Uchino, Yamamoto
Namemedium
2/4 transformation E1010 colony2 1LB(CP)
2/4 transformation E1010 colony2 2LB(CP)

2/8

Miniprep

 Uchino, Egashira, Yamamoto
DNAConcentration/(&microg/ml)260/280260/230
2/7 Liquid culture BAN9123.61.872.07
2/7 Liquid culture BAN9248.81.831.84
2/7 Liquid culture BAN-HIS14268.91.862.15
2/7 Liquid culture BAN-HIS14127.91.852.08
2/7 Liquid culture INP-HIS9354.31.862.24
2/7 Liquid culture INP-HIS9289.41.851.92
2/7 Liquid culture INP-HIS12172.31.872.07
2/7 Liquid culture INP-HIS12286.51.86

2/8

Electrophoresis

 Uchino, Egashira, Yamamoto
Lanesample/(&microl)Length(bp)
11kb ladder /5
22/8 miniprep BAN9 /5
32/8 miniprep BAN9 /5
42/8 miniprep BAN-HIS14 /5
52/8 miniprep BAN-HIS14 /5
62/8 miniprep INP-HIS9 /5
72/8 miniprep INP-HIS9 /5
82/8 miniprep INP-HIS14 /5
92/8 miniprep INP-HIS9 /5

2/9

Miniprep

 Matsumoto, Notsu, Egashira, Yamamoto, Yamada
DNAConcentration/(&microg/ml)260/280260/230
2/8 liquid culture E1010 colony2 1and 258.51.892.16

2/9

Restriction Enzyme Digestion

 Uchino, Egashira, Yamamoto, Yamada
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
2/8 liquid culture E1010 colony2 30010110×M /42240

2/9

Restriction Enzyme Digestion

 Uchino, Yamamoto, Yamada
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
2/8 liquid culture E1010 colony2 5000010×M /211220

2/10

Electrophoresis

 Uchino, Yamamoto, Yamada
Lanesample/(&microl)Length(bp)
11kb ladder /5
22/9 Negative Control E1010 colony2 /102776
32/9 Digestion E1010 colony2 p+x /40732
42/9 Negative Control E0040 colony2 /202799
52/9 Digestion E0040 colony2 p+x /40746
62/9 Negative Control K592009 colony2 /202739
72/9 Digestion K592009 colony2 p+x /40695
82/9 Negative Control E0022 colony2 /202832
92/9 Digestion E0022 colony2 p+x /40

Lane7流出

2/10

Electrophoresis

 Uchino, Yamamoto, Yamada
Lanesample/(&microl)Length(bp)
11kb ladder /5
32/9 NC J23105 colony2 /202105
42/9 Digestion J23105 colony2 E+S /4058
52/9 NC J23100 colony2 /202105
62/9 Digestion J23100 colony2 E+S /4058
72/9 NC B0034 colony3 /202091
82/9 Digestion B0034 colony3 P+S /40

2には何も入れてない

2/10

Miniprep

 Michimori, Egashira, Yamamoto, Yamada
DNAConcentration/(&microg/ml)260/280260/230
E10109.42.130.61
K5920095.21.740.29
E004015.01.940.38
E00228.51.040.86
B003421.21.841.06

E1010　理想値18.5ng/&microLぐらい...
(制限酵素処理100%、ゲル抽出80%として)

2/12

PartsAwaking

 Uchino
DescriptionPartNameBackboneWellPlateYearPlateNumberMilliQ(&microl)
sfGFP-CBDcexK1321342pSB1C318E2015510
sfGFP-CBDclosK1321341pSB1C318C2015510

2/12

Transformation

 Uchino
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
2/12 parts awakening sfGFP-CBDcex1DH5α /20LB(CP)
2/12 parts awakening sfGFP-CBDclos1DH5α /20LB(CP)

2/12

PCR (Target)

 Uchino
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/8 miniprep BAN-His14 4 /0.1VF /0.75m.scFv.Rv /0.75-10.92×KAPA HiFi /12.525
2/8 miniprep BAN-His14 4 /0.1VR /0.75m.scFv.Fw /0.75-10.92×KAPA HiFi /12.525

2/12

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
11kb ladder5
22/12 PCR VF.m.scFv.Rv25
32/12 PCR VR.m.Fw25

2/12

PartsAwaking

 Uchino
DescriptionPartNameBackboneWellPlateYearPlateNumberMilliQ(&microl)
PromoterJ23114J6100219J2015410
PromoterJ23108pSB1C34C2015410

2/12

Transformation

 Uchino, Yamada
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
2/12 Parts awakening J231141DH5α /20LB(AMP)
2/12 Parts awakening J231081DH5α /20LB(AMP)

2/12

Gel Extraction(Measurement)

 Yamada
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
2/12 PCR VF.m.scFv.Rv 18.81.951.52
2/12 PCR VR.m.scFv.Fw 28.62.080.07

2/15

Colony PCR (typeA)

 Egashira
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
2/12 transformation sfGFP-CBDcex colony1VF /0.75 VR /0.75112×KAPA HiFi /12.525
2/12 transformation sfGFP-CBDcex colony2VF /0.75 VR /0.75112×KAPA HiFi /12.525
2/12 transformation sfGFP-CBDclos colony1VF /0.75 VR /0.75112×KAPA HiFi /12.525
2/12 transformation sfGFP-CBDclos colony2VF /0.75 VR /0.75112×KAPA HiFi /12.525

2/15

Electrophoresis

 Kaneko
Lanesample/(&microl)Length(bp)
11kb ladder /5
22/12 transformation sfGFP-CBDcex colony1
32/12 transformation sfGFP-CBDcex colony2
42/12 transformation sfGFP-CBDclos colony1
52/12 transformation sfGFP-CBDclos colony2

2/15

Liquid Culture

 Wan, Yamamoto
Namemedium
2/12 transformation sfGFP-CBDcex colony2LB(CP)
2/12 transformation sfGFP-CBDclos colony2LB(CP)

2/15

Colony PCR (typeA)

 Egashira
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
2/12 transformation J23114 colony1VF 0.75 VR 0.75112×KAPA HiFi /12.525
2/12 transformation J23114 colony2VF 0.75 VR 0.75112×KAPA HiFi /12.525
2/12 transformation J23108 colony1VF 0.75 VR 0.75112×KAPA HiFi /12.525
2/12 transformation J23108 colony2VF 0.75 VR 0.75112×KAPA HiFi /12.525

2/15

PCR (Steps)

 Uchino
StepsTemparatureTimeCycle
Predenature 95℃3min1
Denature 98℃20sec35
Annealing 65℃15sec35
Extension 72℃30sec35
Final Elongation72℃5min1
Final Holding4℃--

2/15

PCR (Target)

 Uchino
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/12 Gel extraction PCR VF/m.scFv.Rv /0.53 + 2/12 Gel extraction PCR VR/m.scFv.Fw /0.35VF /0.75VR /0.75-10.122×KAPA HiFi /12.525
2/12 Gel extraction PCR VF/m.scFv.Rv /0.53 + 2/12 Gel extraction PCR VR/m.scFv.Fw /0.35VF /0.75VR /0.75-10.122×KAPA HiFi /12.525

2/15

Electrophoresis

 Kaneko
Lanesample/(&microl)Length(bp)
11kb ladder /5
2other sample-
3other sample-
4other sample-
5other sample-
6other sample-
7other sample-
8other sample-
9other sample-
10other sample-
11other sample-
12other sample-
13other sample-
142/12 transformation J23114 colony1 /10-
152/12 transformation J23114 colony2 /10
162/12 transformation J23108 colony1 /10
172/12 transformation J23108 colony2 /10

2/15

Gel Extraction

 Yamamoto
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11ka ladder5
22/15 OE-PCR VF/m.scFv.Rv+VR/m.scFv.Fw25
32/15 OE-PCR VF/m.scFv.Rv+VR/m.scFv.Fw25

2/15

Liquid Culture

 Wan, Yamada
Namemedium
2/12 transformation J23114 colony2LB(AMP)
2/12 transformation J23108 colony2LB(AMP)

2/16

Miniprep

 Wan, Yamamoto
DNAConcentration/(&microg/ml)260/280260/230
2/15 liquid culture sfGFP-CBDcex colony243.51.911.60
2/15 liquid culture sfGFP-CBDclos colony274.01.861.82

2/16

PCR (Steps)
StepsTemparatureTimeCycle
Predenature95℃5min1
Denature98℃20sec35
Annealing65℃15sec35
Extension72℃30sec35
Final Elongation72℃5min1
Final Holding4℃--

2/16

PCR (Target)
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/16 miniprep sfGFP-CBDcex colony2 /0.3cex.Fw /0.75VR /0.75-10.72×KAPA HiFi /12.525
2/16 miniprep sfGFP-CBDclos colony2 /0.3cex.Fw /0.75VR /0.75-10.72×KAPA HiFi /12.525

2/16

Gel Extraction(Measurement)

 Yamamoto
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
2/15 OE-PCR VF/m.scFv.Rv+VR/m.scFv.Fw 48.61.830.19

2/16

PCR (Target)

 Yamamoto, Yamada
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/15 Gel Extraction OE-PCR VF/m.scFv.Rv+VR/m.scFv.Fw /0.3VF /0.75VR /0.75-10.72×KAPA HiFi /12.525
2/15 Gel Extraction OE-PCR VF/m.scFv.Rv+VR/m.scFv.Fw /0.3VF /0.75VR /0.75-10.72×KAPA HiFi /12.525

2/16

Miniprep

 Uchino, Yamada
DNAConcentration/(&microg/ml)260/280260/230
2/15 liquid culture J23114178.81.912.20

2/16

PCR (Purification)

 Yamada
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
2/16 PCR m.scFv152.61.862.10

2/16

Restriction Enzyme Digestion

 Matsumoto, Notsu, Wan
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
2/16 PCR m.scFv4100110×H /301130
K59200925100110×H /30030

2/16

Gel Extraction

 Yamamoto
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder5
2m.scFv 2/16 EPcut30
3K592009 2/16EPcut30

2/16

Liquid Culture

 Uchino
Namemedium
2/12 transformation J23108 colony3LB(AMP)

2/17

PCR (Steps)

 Uchino, Kim
StepsTemparatureTimeCycle
Predenature95℃5min1
Denature98℃20sec25
Annealing65℃15sec25
Extension72℃30sec25
Final Elongation72℃5min1
Final Holding4℃--

2/17

PCR (Target)

 Uchino, Kim
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/8 miniprep BAN-His14 /0.1BAN-His-cex.Rv /0.75VF /0.75-10.9KAPA HiFi Ready Mix /12.525
2/8 miniprep BAN-His14 /0.1BAN-His-clos.Rv /0.75VF /0.75-10.9KAPA HiFi Ready Mix /12.525
2/8 miniprep INP-His12 /0.1INP-His-cex.Rv /0.75VF /0.75-10.9KAPA HiFi Ready Mix /12.525
2/8 miniprep INP-His12 /0.1INP-His-clos.Rv /0.75VF /0.75-10.9KAPA HiFi Ready Mix /12.525

2/17

Gel Extraction

 Michimori, Wan, Yamamoto, Kim
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
1blank
22/17PCR BAN-His-CBDcexRv+VF25
32/17PCR BAN-His-CBDclosRv+VF25
42/17PCR INP-His-CBDcexRv+VF25
52/17PCR INP-His-CBDclosRv+VF25
62/16PCR sf-GFP-CBDcex50
72/16PCR sf-GFP-CBDclos50
8blank-
91kb ladder-

2/17

Gel Extraction(Measurement)

 Michimori, Wan, Yamamoto, Kim
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
2/17PCR BAN-His-CBDcexRv+VF10.01.860.43
2/17PCR BAN-His-CBDclosRv+VF34.61.911.81
2/17PCR INP-His-CBDcexRv+VF100.21.901.24
2/17PCR INP-His-CBDclosRv+VF116.51.902.08
2/16PCR sf-GFP-CBDcex244.01.912.08
2/16PCR sf-GFP-CBDclos208.91.902.01

2/17

PCR (Steps)

 Yamamoto
StepsTemparatureTimeCycle
Predenature95℃3min1
Denature98℃20sec35
Annealing65℃15sec35
Extension72℃30sec35
Final Elongation72℃5min1
Final Holding4℃--

2/17

PCR (Target)

 Yamamoto
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/17 Gel Extraction BAN-His-CBDcexRv+VF /8.5 + 3/17 Gel Extraction sf-GFP-CBDcex /0.5VF /0.75VR /0.75-22×KAPA HiFi /12.525
2/17 Gel Extraction BAN-His-CBDclosRv+VF /7 + 3/17 Gel Extraction 2/16PCR sf-GFP-CBDclos /2VF /0.75VR /0.75-22×KAPA HiFi /12.525
2/17 Gel Extraction INP-His-CBDcexRv+VF /6.5 + 3/17 Gel Extraction sf-GFP-CBDcex /1VF /0.75VR /0.75-3.52×KAPA HiFi /12.525
2/17 Gel Extraction INP-His-CBDclosRv+VF /9 + 3/17 Gel Extraction sf-GFP-CBDclos /2VF /0.75VR /0.75-02×KAPA HiFi /12.525

2/17

Miniprep

 Matsumoto, Kim
DNAConcentration/(&microg/ml)260/280260/230
2/16 liquid culture J23108 colony320.81.901.28

2/17

Gel Extraction(Measurement)

 Notsu, Wan
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
2/16 Gel Extraction m.scFv EPcut 14.31.871.09
2/16 Gel Extraction K592009 EPcut14.11.861.60

2/17

Ligation

 Wan, Yamamoto
Vector/(&microl)Insert/(&microl)Ligation High/(&microl)Total/(&microl)
2/16 Gel Extraction K592009 EPcut /2.52/16 Gel Extraction m.scFv EPcut /5.07.515

2/17

Transformation

 Wan, Yamamoto
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
2/17 ligation m.scFv EPcut+K592009 EPcut2DH5α /20LB(CP)

2/18

Gel Extraction(Measurement)

 Kim
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
2/17 OE-PCR BAN-His-CBDcexRv+VF /8.5 + 3/17 Gel Extraction sf-GFP-CBDcex72.71.871.93
2/17 OE-PCR BAN-His-CBDclosRv+VF /7 + 3/17 Gel Extraction 2/16PCR sf-GFP-CBDclos112.11.921.34
2/17 OE-PCR INP-His-CBDcexRv+VF /6.5 + 3/17 Gel Extraction sf-GFP-CBDcex102.41.891.96

2/18

Gel Extraction(Measurement)

 Kim
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
2/17 OE-PCR INP-His-CBDclosRv+VF /9 + 3/17 Gel Extraction sf-GFP-CBDclos103.41.942.04

2/18

Gel Extraction

 Kim
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder8
22/17 OE-PCR BAN-His-CBDcexRv+VF /8.5 + 3/17 Gel Extraction sf-GFP-CBDcex25
32/17 OE-PCR BAN-His-CBDclosRv+VF /7 + 3/17 Gel Extraction 2/16PCR sf-GFP-CBDclos25
42/17 OE-PCR INP-His-CBDcexRv+VF /6.5 + 3/17 Gel Extraction sf-GFP-CBDcex25

2/18

Gel Extraction

 Kim
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder8
22/17 OE-PCR INP-His-CBDclosRv+VF /9 + 3/17 Gel Extraction sf-GFP-CBDclos25
3blank-
4blank-

2/18

Gel Extraction(Measurement)
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
BAN-CBDcex72.31.851.87
BAN-CBDclos79.81.861.26
INP-CBDcex83.61.801.57
INP-CBDclos39.41.761.77

2/18

PCR (Target)

 Kim
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
BAN-CBDcex /0.2VF /1.25VR /1.25-9.8KAPA Pre Mix /12.525
BAN-CBDclos /0.2VF /1.25VR /1.25-9.8KAPA Pre Mix /12.525
INP-CBDcex /0.2VF /1.25VR /1.25-9.8KAPA Pre Mix /12.525
INP-CBDclos /0.2VF /1.25VR /1.25-9.8KAPA Pre Mix /12.525

2/18

Colony PCR (typeA)

 Uchino
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
2/17 transformation m.scFv EPcut+K592009 EPcut colony1VF 1.25 VR 1.259.52×KAPA HiFi Fast Hotstart /12.525
2/17 transformation m.scFv EPcut+K592009 EPcut colony2VF 1.25 VR 1.259.52×KAPA HiFi Fast Hotstart /12.525
2/17 transformation m.scFv EPcut+K592009 EPcut colony3VF 1.25 VR 1.259.52×KAPA HiFi Fast Hotstart /12.525
2/17 transformation m.scFv EPcut+K592009 EPcut colony4VF 1.25 VR 1.259.52×KAPA HiFi Fast Hotstart /12.525

2/18

PCR (Steps)

 Uchino
StepsTemparatureTimeCycle
Predenature95℃2min1
Denature95℃10sec30
Annealing55℃15sec30
Extension72℃30sec30
Final Elongation72℃5min1
Final Holding4℃--

2/18

PCR (Purification)

 Michimori, Kim
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
2/18 PCR m.scFv EPcut+K592009 EPcut colony149.81.920.08
2/18 PCR m.scFv EPcut+K592009 EPcut colony241.11.811.90
2/18 PCR m.scFv EPcut+K592009 EPcut colony347.21.871.99
2/18 PCR m.scFv EPcut+K592009 EPcut colony447.51.871.93

2/19

PCR (Purification)

 Yamamoto
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
BAN-cex/(25)161.11.882.13
BAN-clos/(25)217.91.862.10
INP-cex/(25)158.61.861.98
INP-clos/(25)163.61.85

2/19

Restriction Enzyme Digestion

 Yamamoto
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
BAN-cex12.4100110×H/(3)012.6
BAN-clos9.2100110×H/(3)015.8
INP-cex12.6100110×H/(3)012.4
INP-clos12.2100110×H/(3)012.8

2/16

Gel Extraction

 Kosaka
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
1BAN-cex E, Pcut30
2BAN-clos E, Pcut30
3INP-cex E, Pcut30
01kb ladder5

2/19

Gel Extraction

 Egashira, Wan, Yamamoto
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder5
22/19 Restriction Enzyme INP-clos EPcut30
32/19 Restriction Enzyme E1010 EPcut30
4blank-

2/16

PCR (Purification)

 Wan
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
2/19 Restriction Enzyme BAN-cex E, Pcut9.61.850.84
〃BAN-clos E, Pcut18.91.830.71
〃INP-cex E, Pcut14.91.771.46
〃INP-clos E, Pcut13.41.731.67
〃E1010 E, Pcut19.61.86

2/19

Transformation

 Wan, Egashira, Kim
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
insertDNA/ulVectoeDNA/ulligation hightotal
BAN-cex/4E1010cpsb1c3/2612
BAN-clos/3〃/3612
INP-cex/4.5〃/1.5612
INP-clos/4.5〃/1.56

2/19

Transformation

 Wan
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
2/19 Ligation BAN-cex+E101010DH5a/(20)LB(CP)
〃BAN-clos+E101010DH5a/(20)LB(CP)
〃INP-cex+E101010DH5a/(20)LB(CP)
〃INP-clos+E101010DH5a/(20)

2/19

Restriction Enzyme Digestion

 Michimori, Kim
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
2/18 purification m.scFv EPcut+K592009 EPcut colony112.50.5000.510×H /1.50015
2/18 purification m.scFv EPcut+K592009 EPcut colony212.50.5000.510×H /1.50015
2/18 purification m.scFv EPcut+K592009 EPcut colony312.50.5000.510×H /1.50015
2/18 purification m.scFv EPcut+K592009 EPcut colony412.50.5000.510×H /1.50015

2/19

Electrophoresis

 Egashira
Lanesample/(&microl)Length(bp)
1 1kb ladder /5
22/19 restricion enzyme digestion colony1 /10
32/19 restricion enzyme digestion colony2 /10
42/19 restricion enzyme digestion colony3 /10
52/19 restricion enzyme digestion colony4 /10

2/19

Liquid Culture

 Egashira, Wan
Namemedium
2/17 transformaion m.scFv EPcut+K592009 EPcut colony2LB(CP)
2/17 transformaion m.scFv EPcut+K592009 EPcut colony3LB(CP)
2/17 transformaion m.scFv EPcut+K592009 EPcut colony4LB(CP)

2/19

Colony PCR (typeA)

 Yamada
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
2/19 transformation BAN-His-CBDcex colony1vf 1.25 vr 1.251012.525
〃 colony2〃〃〃〃
〃 colony3〃〃〃〃
2/19 transformation BAN-His-CBDclos colony1〃〃〃〃
〃 colony2〃〃〃〃
〃 colony3〃〃〃〃
2/19 transformation INP-His-CBDcex colony1〃〃〃〃
〃 colony2〃〃〃〃
〃 colony3〃〃〃〃

2/22

Liquid Culture

 Yamada
Namemedium
2/22 PCR BAN-His-cex colony1CP/5ml
〃colony2〃
〃colony3〃
2/22 PCR BAN-His-clos colony3

2/22

Miniprep

 Egashira, Yamada
DNAConcentration/(&microg/ml)260/280260/230
2/19 liquid culture m.scFv EPcut+K592009 EPcut colony2137.41.892.12
2/19 liquid culture m.scFv EPcut+K592009 EPcut colony3164.01.892.09
2/19 liquid culture m.scFv EPcut+K592009 EPcut colony4138.11.862.16

2/19

Ligation

 Wan, Yamamoto
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
insertDNA/ulVectoeDNA/ulligation hightotal
BAN-cex/4E1010cpsb1c3/2612
BAN-clos/3〃/3612
INP-cex/4.5〃/1.5612
INP-clos/4.5〃/1.56

2/23

Transformation

 Wan
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
2/23 Ligation BAN-cex+E10102DH5a/(20)LB(CP)
〃clos〃2〃〃
〃INP-cex〃2〃〃
〃INP-clos〃2〃

2/23

Colony PCR (typeA)

 Egashira
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
2/23 transformation BAN-His-CBDclos colony1vf1.25 vr1.251012.525
〃colony2〃〃〃〃
〃colony3〃〃〃〃
2/23 transformation INP-His-CBDcex colony1〃〃〃〃
〃colony2〃〃〃〃
〃colony3〃〃〃〃
2/23 transformation INP-His-CBDclos colony1〃〃〃

2/24

Miniprep

 Yamada
DNAConcentration/(&microg/ml)260/280260/230
2/22 liqid culture BAN-Hi-cex colony145.31.882.08
〃colony247.11.891.98
〃colony343.81.962.08
2/22 BAN-His-clos colony353.71.82

2/24

Liquid Culture

 Egashira
Namemedium
2/23 transformation BAN-clos colony1LB(CP/5ml)
〃colony2〃
〃colony3〃
2/23 transformation INP-cex colony2〃
〃colony3〃
2/23 transformation INP-clos colony1

2/25

Miniprep

 Egashira, Kim
DNAConcentration/(&microg/ml)260/280260/230
2/23 transformation BAN-His-CBDclos colony179.91.892.15
2/23 transformation BAN-His-CBDclos colony268.01.882.02
2/23 transformation BAN-His-CBDclos colony380.71.942.18
2/23 transformation INP-His-CBDcex colony2106.01.912.17
2/23 transformation INP-His-CBDcex colony3136.71.932.23
2/23 transformation INP-His-CBDclos colony1183.71.912.21

2/25

Sequence

 Michimori
SampleDNA template/(&microl)Primer/(&microl)MilliQ(&microl)Total(&microl)
Bcx 1-F2/24 miniprep BAN-His-CBDcex colony1 /6.6VF /0.23.210
Bcx 1-R2/24 miniprep BAN-His-CBDcex colony1 /6.6VR /0.23.210
Bcx 2-F2/24 miniprep BAN-His-CBDcex colony2 /6.4VF /0.23.410
Bcx 2-R2/24 miniprep BAN-His-CBDcex colony2 /6.4VR /0.23.410
Bcl1-F2/25 miniprep BAN-His-CBDclos colony1 /3.8VF /0.26.010
Bcl1-R2/25 miniprep BAN-His-CBDclos colony1 /3.8VR /0.26.010
Bcl2-F2/25 miniprep BAN-His-CBDclos colony3 /3.7VF /0.26.110
Bcl2-R2/25 miniprep BAN-His-CBDclos colony3 /3.7VR /0.26.110
Icx1-F2/25 miniprep INP-His-CBDcex colony2 /2.8VF /0.27.010
Icx1-R2/25 miniprep INP-His-CBDcex colony2 /2.8VR /0.27.010
Icx2-F2/25 miniprep INP-His-CBDcex colony3 /2.2VF /0.27.610
Icx2-R2/25 miniprep INP-His-CBDcex colony3 /2.2VR /0.27.610
Icl1-F2/25 miniprep INP-His-CBDclos colony1 /1.6VF /0.28.210
Icl1-R2/25 miniprep INP-His-CBDclos colony1 /1.6VR /0.28.210

2/25

Liquid Culture

 Yamada
Namemedium
2/19 transformation BAN-His-cex colony1LB(CP/5ml)
2/19 transformation BAN-His-cex colony2LB(CP/5ml)
2/23 transformation BAN-His-clos colony1LB(CP/5ml)
2/23 transformation BAN-His-clos colony3LB(CP/5ml)
2/23 transformaation INP-His-cex colony2LB(CP/5ml)
2/23 transformation INP-His-cex colony3LB(CP/5ml)
INP-His-clos colony1

2/25

Sequence

 Michimori
SampleDNA template/(&microl)Primer/(&microl)MilliQ(&microl)Total(&microl)
S2-F2/22 miniprep scFv colony2 /2.2VF /0.27.610
S2-R2/22 miniprep scFv colony2 /2.2VR /0.27.610
S3-F2/22 miniprep scFv colony3 /1.8VF /0.28.010
S3-R2/22 miniprep scFv colony3 /1.8VR /0.28.010
S4-F2/22 miniprep scFv colony4 /2.2VF /0.27.610
S4-R2/22 miniprep scFv colony4 /2.2VR /0.27.610

114C-R

2/25

Liquid Culture

 Egashira, Kim, Yamada
Namemedium
2/17 transformaion m.scFv EPcut+K592009 EPcut colony2LB(CP)
2/17 transformaion m.scFv EPcut+K592009 EPcut colony3LB(CP)
2/17 transformaion m.scFv EPcut+K592009 EPcut colony4LB(CP)

2/26

PCR (Target)

 Li
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
J23100VF/ 0.75J23100+RBSRv11PrimeSTARMax Premix/ 12.525
J23108VF/ 0.75J23108+RBSRv11PrimeSTARMax Premix/ 12.525
J23114VF/ 0.75J23114+RBSRv1125

2/26

PCR (Target)

 Li
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
J23100 iGEM PartVF /0.75J23100+RBS Rv /0.75-11PrimeSTAR Max Premix /12.525
J23108 iGEM PartVF /0.75J23108+RBS Rv /0.75-11PrimeSTAR Max Premix /12.525
J23114 iGEM PartVF /0.75J23114+RBS Rv /0.75-11PrimeSTAR Max Premix /12.525

2/26

Electrophoresis

 Egashira
Lanesample/(&microl)Length(bp)
11kb ladder /5-
22/26 PCR J23100 /5193+α
32/26 PCR J23108 /5193+α
42/26 PCR J23114 /5193+α

2/26

PCR (Target)

 Egashira
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
J23100 iGEM Part /0.1VF 0.75J23100+RBS Rv /0.75-10.9KAPA HiFi /12.525
J23108 iGEM Part /0.1VF 0.75J23108+RBS Rv /0.75-10.9KAPA HiFi /12.525
J23114 iGEM Part /0.1VF 0.75J23114+RBS Rv /0.75-10.9KAPA HiFi /12.525

2/26

Gel Extraction

 Egashira
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder 5
22/26 PCR J2310025
32/26 PCR J2310825
42/26 PCR J23114

2/29

Gel Extraction(Measurement)

 Egashira, Wan
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
2/26 PCR J231001.52.710.09
2/26 PCR J231083.618.640.01
2/26 PCR J231141.73.050.13

2/29

PCR (Target)

 Wan, Kim
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
J23100 iGEM Part /0.2VF /1.5J23100+RBS Rv /1.5-21.8KAPA HiFi /2550
J23108 iGEM Part /0.2VF /1.5J23108+RBS Rv /1.5-21.8KAPA HiFi /2550
J23114 iGEM Part /0.2VF /1.5J23114+RBS Rv /1.5-21.8KAPA HiFi /2550

3/1

Electrophoresis

 Yamada
Lanesample/(&microl)Length(bp)
11kb ladder /5-
23/1 PCR J23100 /5about 200
33/1 PCR J23108 /5about 200
43/1 PCR J23114 /5about 200
53/1 PCR m.scFv colony3 /5949

3/1

Electrophoresis

 Kim, Yamada
Lanesample/(&microl)Length(bp)
1100bp ladder /5-
23/1 OE-PCR J23100 + m.scFv colony3 /2about 1200
33/1 OE-PCR J23108 + m.scFv colony3 /2about 1200
43/1 OE-PCR J23114 + m.scFv colony3 /2about 1200

3/1

PCR (Purification)

 Kim, Yamada
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
3/1 OE-PCR J23100 + m.scFv colony3 180.51.891.83
3/1 OE-PCR J23108 + m.scFv colony3183.31.871.96
3/1 OE-PCR J23114 + m.scFv colony3178.81.872.08

3/1

PCR (Target)

 Kim, Yamada
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/1 PCR J23100 /1 + 3/1 PCR m.scFv colony3 /0.5VF /0.5VR /0.75-9.5Prime Star Premix /12.525
3/1 PCR J23108 /1 + 3/1 PCR m.scFv colony3 /0.5VF /0.5VR /0.75-9.5Prime Star Premix /12.525
3/1 PCR J23114 /2 + 3/1 PCR J23114 /0.5VF /0.5VR /0.75-8.5Prime Star Premix /12.525

3/1

Gel Extraction

 Kim, Yamada
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder
23/1 EPcut J23114 + m.scFv20
33/1 EPcut J23108 + m.scFv20
43/1 EPcut J23100 + m.scFv20

3/1

Restriction Enzyme Digestion

 Li, Kim, Yamada
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
3/1 OE-PCR J23100 + m.scFv colony311100110×H /3-1430
3/1 OE-PCR J23108 + m.scFv colony311100110×H /3-1430
3/1 OE-PCR J23114 + m.scFv colony311100110×H /3-1430

3/2

PartsAwaking

 Michimori, Kim, Yamada
DescriptionPartNameBackboneWellPlateYearPlateNumberMilliQ(&microl)
f1 originK314110pSB1C31A2016110
T1 terminator from E. coli rrnBK731722pSB1C31B2016110
Promoter of the CaiF proteinK398326pSB1C31C2016110

3/2

Transformation

 Michimori
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
3/2 Parts Awakening K31411010DH5α /20LB(CP)
3/2 Parts Awakening K73172210DH5α /20LB(CP)
3/2 Parts Awakening K39832610DH5α /20LB(CP)

3/2

Gel Extraction(Measurement)

 Yamada
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
J23100+m.scFv 3/1 EPcut 17.71.901.68
J23108+m.scFv 3/1 EPcut 11.92.040.26
J23114+m.scFv 3/1 EPcut 16.81.971.79

3/2

Restriction Enzyme Digestion

 Wan
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
E002040200210×H /6-1060

3/2

Transformation

 Yamada
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
J23100+pSB1C32DH5α /20LB(CP)
J23108+pSB1C32DH5α /20LB(CP)
J23114+pSB1C32DH5α /20LB(CP)

3/3

PCR (Target)

 Yamada
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/9 miniprep E1010 /1VF 0.75VR 0.75-10Prime Star premix 12.525
2/24 miniprep BAN-His-CBDcex colony2 /1VF 0.75VR 0.75-10Prime Star premix 12.525
2/25 miniprep BAN-His-CBDclos /1VF 0.75VR 0.75-10Prime Star premix 12.525
2/25 miniprep INP-His-CBDcex /1VF 0.75VR 0.75-10Prime Star premix 12.525
2/25 miniprep INP-His-CBDclos /1VF 0.75VR 0.75-10Prime Star premix 12.525

3/3

Gel Extraction(Measurement)

 Egashira
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
3/2 EPcut E002023.11.851.69

3/3

Gel Extraction

 Egashira
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder
23/2 EPcut E0020 (pSB1C3)20

3/3

Ligation

 Wan
Vector/(&microl)Insert/(&microl)Ligation High/(&microl)Total/(&microl)
3/3 Gel Extraction E0020 EPcut /43/2 Gel Extraction J23100+m.scFv EPcut /121632
3/3 Gel Extraction E0020 EPcut /43/2 Gel Extraction J23108+m.scFv EPcut /122040
3/3 Gel Extraction E0020 EPcut /43/2 Gel Extraction J23114+m.scFv EPcut /121632

3/3

Transformation

 Wan, Kim
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
J23100+PSB1C32DH5alpha/ 20LB(CP)
J23108+PSB1C32DH5alpha/ 20LB(CP)
J23114+PSB1C32DH5alphaLB(CP)

3/3

Liquid Culture

 Wan, Kim
Namemedium
K314110 colony1LB(CP)
K731722 colony1LB(CP)
K398326 colony1LB(CP)

3/4

Transformation

 Li, Notsu
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
J23100+pSB1C310DH5α /20LB(CP)
J23108+pSB1C310DH5α /20LB(CP)
J23114+pSB1C310DH5α /20LB(CP)

3/7

Miniprep

 Yamada
DNAConcentration/(&microg/ml)260/280260/230
2/25 liquid culture BAN-His-CBDcex colony271.41.831.43
2/25 liquid culture BAN-His-CBDclos colony161.61.831.90
2/25 liquid culture INP-His-CBDcex colony383.41.941.92
2/25 liquid culture INP-His-CBDclos colony1100.71.871.44

3/7

Transformation

 Wan
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
3/7 miniprep BAN-His-CBDcex2DH5α /20LB(CP)
3/7 miniprep BAN-His-CBDclos2DH5α /20LB(CP)
3/7 miniprep INP-His-CBDcex2DH5α /20LB(CP)
3/7 miniprep INP-His-CBDclos2DH5α /20LB(CP)

3/7

Transformation

 Yamamoto
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
3/3 ligation J23100+pSB1C3DH5α /20LB(CP)
3/3 ligation J23108+pSB1C3DH5α /20LB(CP)
3/3 ligation J23114+pSB1C3DH5α /20LB(CP)

3/7

Colony PCR (typeA)

 Yamamoto
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
3/4 transformtion J23100 colony1VF 1.25 VR 1.2510KAPA2G /12.525
3/4 transformtion J23100 colony2VF 1.25 VR 1.2510KAPA2G /12.525
3/4 transformtion J23108 colony1VF 1.25 VR 1.2510KAPA2G /12.525
3/4 transformtion J23108 colony2VF 1.25 VR 1.2510KAPA2G /12.525
3/4 transformtion J23114 colony1VF 1.25 VR 1.2510KAPA2G /12.525
3/4 transformtion J23114 colony2VF 1.25 VR 1.2510KAPA2G /12.525

3/7

PCR (Steps)

 Yamamoto
StepsTemparatureTimeCycle
Predenature95℃2min1
Denature95℃15sec35
Annealing55℃15sec35
Extension72℃5sec35
Final Elongation72℃10min1
Final Holding4℃--

3/7

Electrophoresis

 Wan, Yamamoto
Lanesample/(&microl)Length(bp)
11kb ladder /5
23/7 colony PCR J23100 colony1 /101000+α
33/7 colony PCR J23100 colony2 /101000+α
43/7 colony PCR J23108 colony1 /101000+α
53/7 colony PCR J23108 colony2 /101000+α
63/7 colony PCR J23114 colony1 /101000+α
73/7 colony PCR J23114 colony2 /101000+α

3/7

PCR (Steps)

 Yamada
StepsTemparatureTimeCycle
Predenature95℃5min1
Denature98℃20sec35
Annealing60℃15sec35
Extension72℃5min35
Final Elongation4℃--
Final Holding4℃--

3/7

PCR (Target)

 Yamada
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/1 purification J23100VF /3VR /3-44KAPA HiFi /50100
3/1 purification J23100VF /3VR /3-44KAPA HiFi /50100
3/1 purification J23108VF /3VR /3-44KAPA HiFi /50100
3/1 purification J23108VF /3VR /3-44KAPA HiFi /50100
3/1 purification J23114VF /3VR /3-44KAPA HiFi /50100
3/1 purification J23114VF /3VR /3-44KAPA HiFi /50100

3/7

Colony PCR (typeA)

 Yamamoto
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
3/7 transformation BAN-CBDcex colony1VF 1.5 VR1.251012.525
3/7 transformation BAN-cex colony2VF 1.5 VR1.251012.525
3/7 transformation BAN-clos colony1VF 1.5 VR1.251012.525
3/7 transformation BAN-closcolony1VF 1.5 VR1.251012.5
3/7 transformation INP-cex colony2VF 1.5 VR1.251012.525
3/7 transformation INP-cex colony2VF 1.5 VR1.251012.525
3/7 transformation INP-clos colony1VF 1.5 VR1.251012.525
3/7 transformation INP-clos colony 2VF 1.5 VR1.251012.525

3/8

PCR (Purification)

 Uchino
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
3/7 PCR J23100423.31.842.20
3/7 PCR J23108439.11.852.11
3/7 PCR J23114451.21.811.91

3/8

Restriction Enzyme Digestion

 Yamamoto
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
3/8 purification J231005100110×H /302030
3/8 purification J231085100110×H /302030
3/8 purification J231145100110×H /302030

3/8

Gel Extraction(Measurement)

 Li, Kim, Yamada
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
J23114 3/8 EPcut13.73.620.03
J23108 3/8 EPcut15.41.831.14
J23100 3/8 EPcut14.51.791.03

3/9

Miniprep

 Kim
DNAConcentration/(&microg/ml)260/280260/230
K73172267.21.701.02
K31411028.91.861.14
K39832695.61.640.94

3/9

Restriction Enzyme Digestion

 Yamamoto
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
3/1 miniprep K31411025100110×H /30030
3/1 miniprep K73172225100110×H /30030
3/1 miniprep K39832625100110×H /30030

3/9

Restriction Enzyme Digestion

 Li
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
3/8 purification J231005100110×H /302030
3/8 purification J231085100110×H /302030
3/8 purification J231145100110×H /302030
3/8 purification J2310020100110×H /30530
3/8 purification J2310820100110×H /30530
3/8 purification J2311420100110×H /30530
30

3/11

Gel Extraction

 Yamada
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder5
23/10 E.P. cut J23100+scFv15
33/10 E.P. cut J23100+scFv15
43/10 E.P.cut J23108+scFv15
53/10 E.P.cut J23108+scFv15
63/10 E.P.cut J23114+scFv15
73/10 E.P.cut J23114+scFv15
83/9 E.P. cut J23100+scFv15
93/9 E.P. cut J23100+scFv15
103/9 Ｅ.P. cut J23119 scFv15
113/9 Ｅ.P. cut J23119 scFv15
123/9 E.P.cut J23114+scFv

3/11

Gel Extraction

 Yamada
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kbp ladder15
23/9 E.P. cut K31411015
33/9 E.P. cut K31411015
43/9 E.P cut K73172215
53/9 E.P cut K73172215
63/9 E.P cut K739832615
73/9 E.P cut K739832615
83/9 E.P> cut J23114+scFv

3/11

PCR (Purification)

 Yoshimoto
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
3/11 Gel Ex J23108+scFv18.91.900.19
3/11 Gel Ex J23108+scFv7.01.920.57
3/11 Gel Ex J23114+scFv5.32.370.25
3/11 Gel Ex J23100+scFv39.21.931.29
3/11 Gel Ex J23108+scFv33.61.820.68
3/11 Gel Ex J23114+scFv27.81.861.25
3/11 Gel Ex pSCIC329.01.86

3/11

Ligation

 Kim
Vector/(&microl)Insert/(&microl)Ligation High/(&microl)Total/(&microl)
3/11 Gel Extraction PSB1C3/ 53/11 Gel Extraction J23100+scfv/ 5.610.621.2
3/11 Gel Extraction PSB1C3/ 53/11 Gel Extraction J23108+scfv/ 511.523
3/11 Gel Extraction PSB1C3/ 53/11 Gel Extraction J23114+scfv/ 512.8

3/11

Transformation

 Kim
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
3/11 Ligation PSB1C3+J23100+scFVDH5α /20LB
3/11 Ligation PSB1C3+J23108+scFvDH5α /20LB
3/11 Ligation PSB1C3+J23114+scFvDH5α /20LB

3/7

Electrophoresis

 Yamada
Lanesample/(&microl)Length(bp)
11kbp ladder
23/8 PCR BANcex colony1515
33/8 PCR BAN-cex colony2515
43/8 PCR BAN-clos colony1487
53/8 PCR BAN-clos-colony2487
63/8 PCR INP-cex colony 11405
73/8 PCR INP-cex colony 21405
83/8 PCR INP-clos colony 11378
93/8 PCR INP-clos colony 21378

3/14

PCR (Target)

 Uchino, Egashira
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
2/25 INPcex colony2VF 0.75 CBDcex ctl. Rv0.7511KAPA HiFi 12.525
2/25 INPcex colony2CBDcex ctl Fw 0.75 VR0.7511KAPA HiFi 12.525
2/18 INPclos Gel ExtractionVF 0.75 CBD clos ctl. Rv0.7511KAPA HiFi 12.525
2/18 INPclos Gel ExtractionCBDclos ctl Fw 0.75 VR0.7511KAPA HiFi 12.525

3/14

Gel Extraction

 Uchino, Egashira
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
43/14PCR INP cex (VF CBD cex ctl.Rv)255
53/14PCR INPcex (CBD cex ctl.Fw VR) 255
63/14PCR INP clos(VF CBDclos ctl.Rv)255
73/14PCR INPclos (CBDclos ctl.Fw VR) 255

3/15

Restriction Enzyme Digestion

 Notsu
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
3/7 miniprep BAN-His-CBDcex colony230114440
3/7miniprep BAN-His-CBDclos colony130114440
3/7miniprep INP-His-CBDcex colony330114440
3/'miniprep INP-His-CBDcloscolony 140115350
2/9 miniprep J23105611070

3/14

OE-PCR (typeA)

 Kim
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
3/14 Gel Ex INP cex 0.5 VR ct Fw + 3/14 Gel Ex INP cex 0.5 VF ct RvVF1.5 VR1.5212550
3/14 Gel Ex INP clos 0.5 VR ct Fw + 3/14 Gel Ex INP clos 0.5 VF ct RvVF1.5 VR1.5212550

3/14

PCR (Steps)

 Kim
StepsTemparatureTimeCycle
Pre Denature95℃5min1
Denature78℃20sec35
Annealing60℃15sec35
Extension72℃45sec35
Final Elongation72℃5min1
Final Holding4℃--

3/14

Colony PCR (typeA)

 Yoshimoto
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
2/8 INP His9VF 0.75 scFv ctl. Rv 0.751112.525
2/8 INP His9scFv ctl Fw 0.75 VR 0.751112.525
2/25 INPcex colony2VF 0.75 CBDcex ctl. Rv0.751112.525
2/25 INPcex colony2CBDcex ctl. Fw 0.75 VR 0.751112.525
2/18 INPclos Gel ExtractionCBDclos ctl. Rv 0.75 VF 0.751112.525
2/18 INPclos Gel ExtractionCBDclos ctl Fw 0.751112.5

3/14

Gel Extraction

 Egashira
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder5
23/14 PCR INP His9 (VF scFv.ctl. Rv)25
33/14 PCR INP His9 (scFv ctl.Fw VR)

3/15

Gel Extraction

 Egashira
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder 5
23/14 OE PCR CBDcex ctl 5010
33/14 OEPCR CBDclos ctl 50

3/15

PCR (Target)

 Egashira
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/25 INPcex colony2VF /1.5CBDcex.ctl.RV /1.5-22KAPA HiFi /2550
2/25 INPcex colony2CBDcex.ctl.Fw /1.5VR /1.5-22KAPA HiFi /2550
2/25 INPclos colony1VF /1.5CBDclos.ctl.Rv /1.5-22KAPA HiFi /2550
2/25 INPclos colony1CBDclos.ctl.Fw /1.5VR /1.5-22KAPA HiFi /2550

3/15

Gel Extraction

 Egashira
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
43/15 PCR INPcex(VF 20, CBDcex ctl.Rv 20)4
53/15 PCR INPcex(CBDcex ctl.Fw20, VR 20)4
63/15 PCR INPclos(VF 20 CBDclos ctl.Rv 20)4
73/15 PCR INPclos(CBDclos ctl.Fw20, VR 20)4

3/15

Liquid Culture

 Li, Uchino
Namemedium
3/11 scFv J23100 colony1,4 colony2,3(plate2)LB(CP) 5×2
3/11 scFv J23108 colony 2(plate1) colony(plate2)LB(CP) 5×2
3/14 scFv J23114 colony 1,2,4(plate1) colony1,2,(plate2)

3/16

PCR (Purification)

 Kim, Yamada
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
3/15 Gelex CBDcex ctl. Fw VR55.31.950.11
3/15 Gelex CBDcex VF ctl.RV8.21.741.04
3/15Gelex CBDclos ctl Fw VR41.21.851.51
3/15Gelex CBDclos VF ctl RV10.61.560.60
3/15Gelex BANcexSP34.91.701.84
3/15Gelex BANclosSP32.01.950.07
3/15 Gelex INPcex SP40.31.761.76
3/15Gelex INPclosSP106.81.830.19

3/16

Gel Extraction

 Egashira
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
33/15 PCR INPcex(VF 20, CBDcex ctl, RV 20)4
43/15 PCR INPclos(VF 20, CBDclos ctl, RV 20)

3/16

Gel Extraction

 Egashira
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
4CBDcex ctl
5CBDclos ctl

3/16

PCR (Target)

 Egashira
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/15 Gel Ex INPcex 0.5(VF ctl.RV) 3/15 Gel Ex INPcex0.2(ctl.Fw VR)VF1.25VR1.25KAPA taq 12.59.325
3/15 GEl EX INPclos 0.5(VF ctl RV) 3/15Gel Ex INPclos0.2(ctl.Fw VR)VF1.25KAPA taq 12.59.325

3/16

PCR (Target)

 Kim
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/14 Gel Extraction INP-His9 VR/ct/Rv /0.5 + 3/14 Gel Extraction INP-His9 /0.5 VF /1.5VR /1.5-212×KAPA HiFi /2550

3/16

PCR

 Egashira
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)KAPA HiFi/(&microl)Total/(&microl)
2/8 INP HIs9VF 1.5 scFvctl. RV1.5222550
2/8 INP His9VR1.5 scFvctl. Fw 1.5 VR1.52225

3/16

Miniprep

 Egashira, Yamamoto
DNAConcentration/(&microg/ml)260/280260/230
J23100 scFv c1-p1135.71.851.76
J23100 scFv C4-p1135.11.861.85
J23100 scFv c3-p2155.01.872.04
J23100 scFv c3-p2205.91.761.32
J23108 scFv c2-p1166.51.882.06
J23108 scFvc1-p295.71.841.83
J23114 scFv c1-p1145.71.801.99
J23114 scFv c2-p1111.71.902.16
J23114 scFv c4-p1169.21.902.11
J23114 scFv c1-p2127.41.872.00
J23114 scFv c2-p2136.21.902.13

3/16

Sequence

 Michimori
SampleDNA template/(&microl)Primer/(&microl)MilliQ(&microl)Total(&microl)
100A-F3/15 miniprep J23100 scFv c1-p1 /2.2VF /0.27.610
100A-R3/15 miniprep J23100 scFv c1-p1 /2.2VR /0.27.610
100B-F3/15 miniprep J23100 scFv c4-p1 /2.2VF /0.27.610
100b-R3/15 miniprep J23100 scFv c4-p1 /2.2VR /0.27.610
100C-F3/15 miniprep J23100 scFv c2-p2 /1.9VF /0.27.910
100C-R3/15 miniprep J23100 scFv c2-p2 /1.9VR /0.27.910
100D-F3/15 miniprep J23100 scFv c3-p2 /1.5VF /0.28.310
100D-R3/15 miniprep J23100 scFv c3-p2 /1.5VR /0.28.310
108A-F3/15 miniprep J23108 scFv c2-p1 /1.8VF /0.28.010
108A-R3/15 miniprep J23108 scFv c2-p1 /1.8VR /0.28.010
108B-F3/15 miniprep J23108 scFv c1-p2 /3.1VF /0.26.710
108B-R3/15 miniprep J23108 scFv c1-p2 /3.1VR /0.26.710
114A-F3/15 miniprep J23114 scFv c1-p1 /2.1VF /0.27.710
114A-R3/15 miniprep J23114 scFv c1-p1 /2.1VR /0.27.710
114B-F3/15 miniprep J23114 scFv c2-p1 /2.7VF /0.27.110
114B-R3/15 miniprep J23114 scFv c2-p1 /2.7VR /0.27.110

114C-R

3/16

Sequence

 Michimori
SampleDNA template/(&microl)Primer/(&microl)MilliQ(&microl)Total(&microl)
114C-F3/15 miniprep J23114 scFv c4-p1 /1.8VF /0.28.010
114C-R3/15 miniprep J23114 scFv c4-p1 /1.8VR /0.28.010
114C-F3/15 miniprep J23114 scFv c1-p2 /2.4VF /0.27.410
114C-R3/15 miniprep J23114 scFv c1-p2 /2.4VR /0.27.410
114C-F3/15 miniprep J23114 scFv c2-p2 /2.2VF /0.27.610
114C-R3/15 miniprep J23114 scFv c2-p2 /2.2VR /0.27.610

114C-R

3/16

Gel Extraction

 Egashira
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
23/15 PCR INP His9(VF scFv CH. Rv)20
33/15 PCR INP His9 (scFv ctl, Fw VR)

3/16

Gel Extraction

 Egashira
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder 5
23/15 PCR INP His9 (VF scFv ctl RV)

3/16

Gel Extraction

 Egashira
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder
2scFv (VF ctl.Rv)
3

3/16

OEPCR (Target)

 Egashira
Sample1/(&microl)Sample2/(&microl)Primer/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/15 Gel Extraction scfv (VF ctl.Rv)/ 0.53/15 Gel Extraction scfv (ctl.Fw VR)VF/ 1.25, VR/1.259.3KAPA Taq/ 12.525

3/16

PCR

 Egashira
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
scFv (VF ctl.RV) 0.5VF 1.25 ctl.Rv) 1.259.512.525

3/16

PCR (Target)

 Egashira
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/16 Gel Extraction scFv(VF.ctl.Rv) /0.25+3/16 Gel Extraction scFv(ctl.Fw.VR) /0.25VF /1.25VR /1.25-9.52×KAPA HiFi /12.525

3/17

PCR (Target)

 Egashira
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/16 Gel Ex scFv(VF ctl.Rv)0.25, 3/15 Gel EX CBDcex(ctl.Fw VR) 0.25VF 1.25VR1.25KAPA Taq 12.59.525
3/16 Gel Ex scFv(VF ctl.Rv) 0.25, Gel Ex CBDclos(ctl,Fw VR) 0.25VF 1.25VR1.25KAPA Taq 12.525

3/17

Electrophoresis

 Egashira
Lanesample/(&microl)Length(bp)
33/16 OEPCR CBDcex ctl 25500
43/16 OEPCR CBDclos ctl 25500

3/17

Electrophoresis

 Egashira
Lanesample/(&microl)Length(bp)
2CBDcex ctl 25
3CBDclos ctl 25
41kb ladder 5

3/17

PCR (Target)

 Egashira
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/16 Gel Ex scFv(VF ctl,Rv)0.1 3/15 Gel Ex CBDcex(ctl,Fw VR) 0.1 3/15 Gel Ex CBDcex(ctl,Fw VR) 0.1VF 1.25VR1.25KAPA 12.59.825
3/16 Gel Ex scFv(VF ctl,Rv) 0.1, 3/15 GelEx CBDclos(ctl,Fw VR) 0.1 ctl,Rv)0.1 3/15 Gel Ex CBDcex(ctl,Fw VR) 0.1VF 1.25VR1.25KAPA 12.59.825

3/17

Miniprep

 Yamamoto
DNAConcentration/(&microg/ml)260/280260/230
3/8 Liquid Culture BAN-His-cex39.31.911.67
3/8 Liquid Culture BAN-His=clos45.11.841.44

3/17

PCR (Steps)

 Yamamoto
StepsTemparatureTimeCycle
Pre Denature95℃3min1
Denature95℃15sec40
Annealing55℃15sec40
Elongation72℃1min40
Final Elongation72℃1min1
Final Holding4℃--

3/17

PCR (Target)

 Yamamoto
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/17 miniprep BAN-His-cexCBDcexFw 1.25VR1.2512.5925
3/17 miniprep BAN-His-closCBDclosFw 1.25VR1.2512.5925
2/9 miniprep J23105_1VF 1.25RFP-cex RV 1.2512.5925
2/9 miniprep J23105_1VF 1.25RFP-clos Rv 1.2512.5925

3/17

Electrophoresis

 Egashira
Lanesample/(&microl)Length(bp)
11kb ladder 5
2

3/17

PCR (Target)

 Egashira
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/16 Gel Extraction scFv(VF.ctl.Rv) /0.1+3/15 Gel Extraction scFv(ctl.Fw.VR) /0.1VF /1.25VR /1.25-9.82×KAPA HiFi /12.525

3/18

Gel Extraction

 Yamamoto
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder 5
23/10 PCR BAN-cex CBDcexFW VR 25
33/10 PCR BAN-clos CBDclosFw VR 25
43/10 PCR J23105 VF RFPcexRv 25
53/10 PCRJ23105 VF RFPclos 25

3/18

Gel Extraction(Measurement)

 Yamamoto
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
3/17 PCR BAN-CBDcex CBDcex.Fw+Rv127.71.901.57
3/17 PCR BAN-CBDclos CBDclos.Fw+VR93.41.922.19
3/17 PCR J23105 VF+RFP-CBDcex.Rv104.01.841.50
3/17 PCR J23105 VF+RFP-CBDclos.Rv123.01.871.84

3/18

PCR (Steps)

 Yamamoto
StepsTemparatureTimeCycle
Pre Denature95℃3min1
Denature95℃15sec40
Annealing60℃15sec40
Elongation72℃1min40
Final Elongation72℃1min1
Final Holding4℃--

3/18

PCR (Target)

 Yamamoto
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/18 Gel Extraction BAN-His-cex 0.1 cexFw+VR , 3/18Gel Extraction J23105 VF+RFP-cexRvVF 1.25VR 1.25KAPATaq 12.59.825
3/18 Gel Extraction BAN-His-clos 0.1 closFw+VR , 3/18Gel Extraction J23105 VF+RFP-closRvVF 1.25VR 1.25KAPATaq 12.59.825

3/18

Gel Extraction

 Yamamoto
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder
23/18 OEPCR RFP-cex
33/18 OEPCR RFP-clos

3/18

PCR (Target)

 Egashira
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
2/25 INP cex colony2 0.1×2VF 0.75×2 ctl RV 0.7510.9×212.5×225×2
2/25 INP cex colony2 0.1×2ctl Fw 0.75×2 VR 0.75×210.9×212.5×225×2
2/25 INP cex colony2 0.1×2VF 0.75×2 VR 0.75×210.9×212.5×225×2
2/25 INP cex colony2 0.1×2nothing12.412.5×225×2
2/25 INP clos colony1 0.1VF ctl.Rv 0.75×210.912.5×225×2
2/25 INP clos colony1 0.1ctl.Rv 0.75×210.912.5×225×2
2/25 INP clos colony1 0.1VF 0.75×2 VR 0.75×210.912.5×225×2
2/25 INP clos colony1 0.1nothing12.412.5×225×2

3/18

Colony PCR (typeA)

 Notsu
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
INP-His=CNDcex+RFP colony 1(3/16 ligation product)VF 1.25, VR .1251012.525
INP-His-CBDcex+RFP colony (3/16 ligation product)VF 1.25, VR .1251012.525
INP-His-CBDclos+RFP colony1(3/16 ligation product)VF 1.25, VR .1251012.525
INP-His-CBDclos+RFP colony 2(3/16 ligation product)VF 1.25, VR .1251012.525
INP-His-CBDcex+RFP colony1( 3/16 ligation product)VF 1.25, VR .1251012.525
BAN-His-CBDcex+RFP colony2 (3/16 ligation product)VF 1.25, VR .1251012.525
BAN-His-CBDclos+RFP colony1(3/16 ligation product)VF 1.25, VR .1251012.525
BAN-His-CBDclos+RFP colony2(3/16 ligation product)VF 1.25, VR .1251012.525

3/18

Electrophoresis

 Kim, Yamada
Lanesample/(&microl)Length(bp)
11kb ladder 5
21 kb ladder 5
32/25 INPcex colony2 10
42/25 INPcex colony2 10
52/25 INPcex colony2 10
62/25 INPcex colony210
72/25 INPclos colony1 10
82/25 INPclos colony1 10
92/25 INPclos colony1 10
102/25 INPclos colony1 10
11100bp ladder 5
121kbp ladder 5

3/18

PCR

 Egashira
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
INP HIs9 0.1x2VF 0.75x2 ctl. Rv 0.75x210.9x212.5x225x2
INP HIs9 0.1x2ctl.Fw 1.75 VR 0.7510.9x212.5x225x2
INP HIs9 0.1x2VF 0.75 VR 0.7510.9x212.5x225x2
INP HIs9 0.1x2nothing12.412.5x2

3/22

Electrophoresis

 Notsu
Lanesample/(&microl)Length(bp)
11kb ladder /7
2INP-His-CBDcex+RFP colony1 (3/19 PCR) /20
3INP-His-CBDcex+RFP colony2 (3/19 PCR) /20
4blank
5INP-His-CBDclos+RFP colony1 (3/19 PCR) /20
6INP-His-CBDclos+RFP colony2 (3/19 PCR) /20
7blank
8BAN-His-CBDcex+RFP colony1 (3/19 PCR) /20
9BAN-His-CBDcex+RFP colony2 (3/19 PCR) /20
10blank
11BAN-His-CBDclos+RFP colony1 (3/19 PCR) /20
12BAN-His-CBDclos+RFP colony2 (3/19 PCR) /20
13blank
14100bp ladder /7
15-
16-
17-

3/22

Electrophoresis

 Egashira
Lanesample/(&microl)Length(bp)
72/25 INPcexcolony2 (5) 10
82/25 INPcexcolony2 (6) 10
92/25 INPcexcolony2 (7) 10
102/25 INPcexcolony2 (8) 10
112/25 INPclos colony1(9) 10
122/25 INPclos colony1(10) 10
132/25 INPclos colony1(11) 10
142/25 INPclos colony1(12) 10
16100b ladder 5
171kb ladder 5

3/22

Electrophoresis

 Egashira
Lanesample/(&microl)Length(bp)
53/22 PCR CBDcex(VF/ctl.Rv) 10
63/22 PCR CBDcex(ctl.Fw/VR) 10
73/22 PCR CBDclos(VP/ctl.RV) 10
83/22 PCR CBDclos(ctl.Fw/VR) 10

3/22

PCR (Target)
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
2/25 INPcex colony2 0.1VF 1.5 ctl, Rv 1.521.92550
2/25 INPcex colony2 0.1ctl.Fw 1.5 ,VR 1.521.92550
2/25 INPclos colony1 0.1VF 1.5, ctl,Rv 1.521.92550
2/25 INPclos colony1 0.1ctl,Fw 1.5 VR 1.521.92550

3/22

Miniprep

 Notsu, Yamada
DNAConcentration/(&microg/ml)260/280260/230
BANcex+RFP col.1147.81.892.18
BANcex+RFP col.2 133.91.872.18
BANcex+RFP col.1132.41.852.19
BANclos+RFP col.2126.41.852.22
INPcex+RFP col.1179.31.872.18
INPcex+RFP col.2144.91.872.19
INPclos+RFPcol.1152.21.872.23
INPclos+RFPcol.2152.41.86

3/22

PCR (Target)

 Yamada
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/8 INP-His9 0.1VF 1.5ctl.Rv 1.52521.950
2/8 INP-His9 0.1scFv ctl.Fw 1.5VR 1.52521.9

3/22

Miniprep

 Egashira, Kim
DNAConcentration/(&microg/ml)260/280260/230
3/22 PCR scFv (VF/ctl.RN)105.41.811.54
3/22 PCR scFv (ctl. FV/VR)127.41.87

3/22

OEPCR (Target)

 Egashira
Sample1/(&microl)Sample2/(&microl)Primer/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/23 Gel Extraction scfv (VF ctl.Rv)/ 0.13/23 Gel Extraction scfv (ctl.Fw VR)/ 0.1VF/ 1.25, VR/1.2510.8KAPA Taq/ 12.525

3/23

Electrophoresis

 Egashira
Lanesample/(&microl)Length(bp)
53/22 PCR CBDcex(VF/ctl.Rv) 40
63/22 PCR CBDcex(ctl.Fw/VR) 40
73/22 PCR CBDclos(VF/ctl.Rv) 40
83/22 PCR CBDclos(ctl.Fw/VR) 40

3/23

PCR (Steps)

 Kim
StepsTemparatureTimeCycle
Pre Denature95℃2min1
Denature98℃20sec35
Annealing60℃15sec35
Elongation72℃15sec35
Final Elongation72℃5min1
Final Holding10℃--

3/23

PCR (Target)

 Kim
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/18 Gelex BANcex Fw+VR 0.1, 3?18 Gelex J23105 VF+cexRv 0.1VF1.25 VR 1.2512.59.825
3/18 Gelex BANclos closFV VR 0.1, 3/18Gelex J23105 VF+closR 0.1VF 1.25 VR 1.2512.59.825

3/23

Gel Extraction (Measurement)

 Egashira
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
3/22 PCR CBDcex(ctlFw/VR)26.91.751.04
3/22PCR CBDclos(ctl Fw/VR)100.61.901.65

3/23

Electrophoresis

 Kim
Lanesample/(&microl)Length(bp)
33/18 OEPCR BANcex cexFw+VR J23105 VF+cexRv 25
4BANclos clos Fw+VR J23105 VF+clos 25
11kb ladder 10
2100bp ladder 10

3/23

Electrophoresis

 Egashira
Lanesample/(&microl)Length(bp)
43/23 OEPCR CBDcex ctl 25
53/23 OEPCR CBDclos ctl 25

3/23

PCR (Target)

 Egashira
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
3/23 Gel Ex scFv(VF/ctl.Rv) 0.1, 3/23 Gel Ex CBDcex(ctl.Fw/VR) 0.1VF 0.75 VR 0.7510.812.525
3/23 Gel Ex scFv (VF/ctl.Rv), 3/23 Gel Ex clos(ctl.Fw/VR)VF 0.75 VR 0.7510.812.525

3/23

Electrophoresis

 Yamada
Lanesample/(&microl)Length(bp)
11kb ladder 5
2100b ladder 7.5
33/22 PCR scFv(VP/ctl.Rv) 40
43/22 PCR scFv(ctl.Fw/VR)

3/24

PCR (Steps)

 Kim
StepsTemparatureTimeCycle
Pre Denature95℃2min1
Denature98℃20sec30
Annealing60℃15sec30
Elongation72℃15sec30
Final Elongation72℃5min1
Final Holding10℃--

3/24

PCR (Target)

 Kim
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/18 Gelcex BAN cex cexFv VR 0.2, 3/18 Gelex J23105 VF cexFv 0.2VF1.25VR 1.2512.59.625
3/18 Gelcex BAN clos clos Fv VR 0.2, 3/18 Gelex J23105 VF clos Rv 0.2VF 1.25VR 1.2512.59.625

3/24

PCR (Purification)

 Egashira
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
3/24 OEPCR CBDcex ctl468.61.822.06
3/24 OEPCR CBDclos ctl321.81.860.45

3/24

PCR (Purification)

 Kim
DNAConcentration/(&microg/ml)260/280260/230
3/24 OEPCR BANcex ctl77.21.811.40
3/24 OEPCR BAN clos ctl23.81.751.56

3/24

PCR (Purification)

 Kim
DNAConcentration/(&microg/ml)260/280260/230
3/24 OEPCR BANcex ctl77.21.811.40
3/24 OEPCR BAN clos ctl23.81.751.56

3/24

PCR (Purification)

 Egashira
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
3/24 OEPCR scFv ctl394.21.83

3/24

Restriction Enzyme Digestion

 Egashira
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
3/24 OE PCR scFv ctl 5.111319.930

3/24

Transformation

 Egashira
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
PCBTC3+scFv+ctl.DH5α

3/25

Restriction Enzyme Digestion

 Egashira
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
3/24 OEPCR CBDcex ctl4.3100110×H /3020.730
3/24 OEPCR CBDclos ctl6.2100110×H /3018.830
3/15 J23100+scFv C3-P29.7100110×H /3015.330

3/25

Restriction Enzyme Digestion

 Kim
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
3/24 OEPCR BAN-CBDcex ctl10100110×H /301530
3/24 OEPCR BAN-CBDclos ctl10100110×H /301530

3/25

Ligation

 Yamada
Vector/(&microl)Insert/(&microl)Ligation High/(&microl)Total/(&microl)
3/24 EPcut pSB1C3 /83/24 EPcut BAN-CBDcex.ctl /3.711.723.4
3/24 EPcut pSB1C3 /73/24 EPcut BAN-CBDclos.ctl /4.711.723.4

3/25

Transformation

 Yamada
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
pSB1C3+BAN-CBDcex.ctl 3/25 ligation-20LB(CP)
pSB1C3+BAN-CBDclos.ctl 3/25 ligation-20LB(CP)
pSB1C3+CBDcex 3/24 ligation-20LB(CP)
pSB1C3+CBDclos 3/24 ligation-20LB(CP)
pSB1C3+scFv.ctl 3/24 ligation-20LB(CP)

3/28

Colony PCR (typeA)

 Yamada
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
BANcex ctl col.1VF 1.25 VR 1.251012.525
BAncex ctl col.2VF 1.25 VR 1.251012.525
BANclos ctl col.1VF 1.25 VR 1.251012.525
BANclos ctl col.2VF 1.25 VR 1.251012.525
CBDcex col.1VF 1.25 VR 1.251012.525
CBDcex col.2VF 1.25 VR 1.251012.525
CBDclos col,1VF 1.25 VR 1.251012.525
CBDclos col.2VF 1.25 VR 1.251012.5
scFv ctl col.1
scFv ctl col.2

3/28

Electrophoresis
Lanesample/(&microl)Length(bp)
11kb ladder(10)
2100bb ladder(10)
3BANcex ctl col.1
4BANcex ctl col.2
5BANclos ctl col.1
6BANclos ctl col.2
7CBDcex col.1
8CBDcex col.2
9CBDclos col.1
10CBDclos col.2
11scFv ctl col.1
12scFv ctl col.2

3/1

Electrophoresis

 Li, Kim
Lanesample/(&microl)Length(bp)
1100bp ladder /6
2,3J23114 /50 193+α
4,5J23100 /50193+α
6,7J23108 /50193+α

3/1

PCR (Purification)

 Yamamoto, Kim
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
2/29 PCR J231144.71.590.01
2/29 PCR J231002.42.600.01
2/29 PCR J231083.84.510.01

3/1

PCR (Target)

 Yamada
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
3/1 PCR purification J23100 /5VF /0.75J23100Rv /0.75-6Prime Star Premix /12.525
3/1 PCR purification J23108 /5VF /0.75J23108Rv /0.75-6Prime Star Premix /12.525
3/1 PCR purification J23114 /5VF /0.75J23114Rv /0.75-6Prime Star Premix /12.525
2/22 miniprep m.scFv colony3 /0.5BAN+scFv /0.75VR /0.75-10.5Prime Star Premix /12.525

7/22

Liquid Culture

 Yamamoto
Namemedium
7/21 transformation BclA-His-scFv colony1LB(CP)
7/21 transformation BclA-His-scFv colony2LB(CP)
7/21 transformation INP-His-scFv colony1LB(CP)
7/21 transformation INP-His-scFv colony2LB(CP)
7/21 transformation BclA-His-CBDcex colony1LB(CP)
7/21 transformation BclA-His-CBDcex colony2LB(CP)
7/21 transformation BclA-His-CBDclos colony1LB(CP)
7/21 transformation BclA-His-CBDclos colony2LB(CP)
7/21 transformation INP-His-CBDcex colony1LB(CP)
7/21 transformation INP-His-CBDcex colony2LB(CP)
7/21 transformation INP-His-CBDclos colony1LB(CP)
7/21 transformation INP-His-CBDclos colony 2LB(CP)

7/25

Miniprep

 Yamamoto
DNAConcentration/(&microg/ml)260/280260/230
7/22 liquid culture BclA-His-scFv1183.51.852.05
7/22 liquid culture BclA-His-scFv2379.01.862.20
7/22 liquid culture INP-His-scFv1189.61.852.21
7/22 liquid culture INP-His-scFv2375.31.852.21
7/22 liquid culture BclA-His-CBDcex191.91.871.90
7/22 liquid culture BclA-His-CBDcex2187.11.871.98
7/22 liquid culture BclA-His-CBDclos141.21.882.21
7/22 liquid culture BclA-His-CBDclos2109.41.942.24
7/22 liquid culture INP-His-CBDcex171.01.731.19
7/22 liquid culture INP-His-CBDcex255.51.892.23
7/22 liquid culture INP-His-CBDclos188.61.821.61
7/22 liquid culture INP-His-CBDclos275.81.862.17

7/25

Restriction Enzyme Digestion

 Yamamoto
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
2/25 miniprep BclA-His-scFv110.9000010×L /3014.130
2/25 miniprep BclA-His-scFv25.3010110×H /33.016.730
2/25 miniprep INP-His-scFv110.5000010×L /3014.530
2/25 miniprep BclA-His-CBDcex210.7001110×M /3014.330
2/25 miniprep BclA-His-CBDclos218.3001110×M /306.730
2/25 miniprep INP-His-CBDcex125001110×M /30030
2/25 miniprep INP-His-CBDclos122.6001110×M /302.430

Samples of "2/25 miniprep BclA-His-scFv1" and "2/25 miniprep INP-His-scFv1" include 2&microl of DraⅡ(Eco0109Ⅰ) (DraⅡis restriction enzyme).

7/26

Gel Extraction

 Yamamoto
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder 5
27/25 restriction enzyme digestion BclA-His-scFv1 DraⅡcut30
37/25 restriction enzyme digestion BclA-His-scFv2 XPcut30
47/25 restriction enzyme digestion INP-His-scFv1 DraⅡcut30
57/25 restriction enzyme digestion BclA-His-CBDcex2 SPcut30
67/25 restriction enzyme digestion BclA-His-CBDclos2 SPcut30
77/25 restriction enzyme digestion INP-His-CBDcex1 SPcut30
87/25 restriction enzyme digestion INP-His-CBDclos1 SPcut30

7/26

Gel Extraction(Measurement)

 Yamamoto
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
7/26 Gel Extraction BclA-His-scFv2 XPcut3.85.140.01
7/26 Gel Extraction BclA-His-CBDcex2 SPcut10.43.240.01
7/26 Gel Extraction BclA-His-CBDclos2 SPcut6.02.670.01
7/26 Gel Extraction INP-His-CBDcex1 SPcut8.12.030.02
7/26 Gel Extraction INP-His-CBDclos1 SPcut14.62.570.02

7/26

Restriction Enzyme Digestion

 Yamamoto
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
7/25 miniprep BclA-His-scFv110.9000010×L /3014.130
7/25 miniprep BclA-His-scFv13.6100110×H /303.410
7/25 miniprep BclA-His-scFv13.60000006.410
7/25 miniprep BclA-His-scFv25.3010110×H /33.016.730
/25 miniprep INP-His-scFv110.5000010×L /3014.530
/25 miniprep INP-His-scFv13.5000110×H /103.510
/25 miniprep INP-His-scFv13.50000006.510
7/25 miniprep BclA-His-CBDcex210.7001110×M /3014.330
7/25 miniprep BclA-His-CBDclos218.3001110×M /306.730
7/25 miniprep INP-His-CBDcex225001110×M /30030
7/25 miniprep INP-His-CBDclos225001110×M /30030

Samples of "2/25 miniprep BclA-His-scFv(10.9&microl)" and "2/25 miniprep INP-His-scFv1(10.5&microl)" include 2&microl of DraⅡ(Eco0109Ⅰ) (DraⅡis restriction enzyme).

7/26

Gel Extraction

 Yamamoto
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder5
22/26Restriction Enzyme Digestion BclA-His-scFv1 DraⅡcut30
32/26Restriction Enzyme Digestion BclA-His-scFv1 XPcut10
42/26Restriction Enzyme Digestion BclA-His-scFv110
52/26Restriction Enzyme Digestion INP-His-scFv1 DraⅡcut30
62/26Restriction Enzyme Digestion INP-His-scFv1EPcut10
72/26Restriction Enzyme Digestion INP-His-scFv1 10
---
1'1kb ladder5
2'2/26Restriction Enzyme Digestion BclA-His-scFv2 XPcut30
3'2/26Restriction Enzyme Digestion BclA-His-CBDcex2 SPcut30
4'2/26Restriction Enzyme Digestion BclA-His-CBDclos2 SPcut30
5'2/26Restriction Enzyme Digestion INP-His-CBDcex2 SPcut30
6'2/26Restriction Enzyme Digestion INP-His-CBDclos2 SPcut30

7/27

Ligation

 Yamamoto
Insert DNA(7/27purification) /&micro?Vector DNA(7/25purification) /&micro?Ligation High /&micro?Total
-BclA-scFv1 DraⅡcut /7.57.515
-INP-scFv1 DraⅡcut /7.57.515
BclAscFv2 XPcut /7.16BclA-CBDcex2 SPcut /0.347.515
BclAscFv2 XPcut /3.75BclA-CBDclos2 SPcut /3.757.515
BclAscFv2 XPcut /1.88INP-CBDcex2 SPcut /5.627.515
BclAscFv2 XPcut /5.0INP-CBDclos2 SPcut /2.57.515

7/27

Transformation

 Yamamoto
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
BclAscFv1 DraⅡcut2DH5α /20LB(CP)
INPscFv1 DraⅡcut2DH5α /20LB(CP)
BclAscFv+BclACBDcex2DH5α /20LB(CP)
BclAscFv+BclACBDclos2DH5α /20LB(CP)
BclAscFv+INPCBDcex2DH5α /20LB(CP)
BclAscfv+INPCBDclos2DH5α /20LB(CP)

7/27

Gel Extraction(Measurement)

 Yamamoto
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
7/27 Gel Extraction BclA-His-scFv1 DraⅡcut4.8-7.111.04
7/27 Gel Extraction INP-His-scFv1 DraⅡcut40.41.321.00
7/27 Gel Extraction BclA-His-scFv2 XPcut9.3-4.240.02
7/27 Gel Extraction BclA-His-CBDcex2 SPcut174.51.320.84
7/27 Gel Extraction BclA-His-CBDclos2 SPcut9.75.840.19
7/27 Gel Extraction INP-His-CBDcex2 SPcut3.7-2.820.14
7/27 Gel Extraction INP-His-CBDclos2 SPcut22.62.510.21

7/28

Restriction Enzyme Digestion

 Yamamoto
DNA /&micro?EcoR1 /&micro?Xba1 /&micro?Spe1 /&micro?Pst1 /&micro?DraⅡ /&micro?Buffer /&micro?BSA /&micro?MilliQ /&micro?Total /&micro?
BclAscFv2 /5.30000110×L /3020.730
BclAscFv2 /5.30101010×H /3316.730
INPscFv2 /5.30000110×L /3020.730
BclAcex2 /10.70011010×M /3014.330
BclAclos1 /250011010×M /30030
INPcex1,2 /250011010×M /30030
INPclos1,2 /250011010×M /30030

7/28

Gel Extraction

 Kaneko
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
7/28 BclA-scFv2 XPcut

0.5&microg/&microl
260/280 2.36
260/230 1.15

7/28

Transformation

 Kaneko
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
2/22 scFv colony2 2DH5α /20

7/28

Ligation

 Yamamoto
Insert DNA /(&microl) Vector DNA /(&microl) Ligation high /(&microl) Total /(&microl)
- 7/28 Restriction Enzyme BclA-scFv DraⅡcut /7.5 7.5 15
- 7/28 Restriction Enzyme INP-scFv DraⅡcut /7.5 7.5 1.5

7/28

Transformation

 Yamada
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
7/28 ligation BclA-scFv control2DH5α /20LB(CP)
7/28 ligation INP-scFv control2DH5α /20LB(CP)
2/22 miniprep BAN-scFvDH5α /20

7/28

Liquid Culture

 Yamamoto
Namemedium
BclA-scFv glycerol stock LB(CP)
7/21 transformation INP-scFv colony1LB(CP)
7/21 transformation BclA-CBDcex colony1LB(CP)
7/21 transformation BclA-CBDclos colony1LB(CP)
7/21 transformation INP-CBDcex colony1LB(CP)
7/21 transformation INP-CBDclos colony1LB(CP)
7/21 transformation LB(CP)

7/21

Liquid Culture

 Yamamoto
Namemedium
BclA-His-scFv glycerol stock1LB(CP)
BclA-His-scFv glycerol stock2LB(CP)
BclA-His-CBDcex glycerol stock1LB(CP)
BclA-His-CBDcex glycerol stock2LB(CP)
BclA-His-CBDclos glycerol stock1LB(CP)
BclA-His-CBDclos glycerol stock2LB(CP)
INP-His-CBDcex glycerol stock1LB(CP)
INP-His-CBDcex glycerol stock2LB(CP)
INP-His-CBDclos glycerol stock1LB(CP)
INP-His-CBDclos glycerol stock2LB(CP)

7/21

Transformation

 Yamamoto
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
2/8 miniprep INP-His-scFv1DH5α /20LB(CP)
2/8 miniprep BclA-His-scFv141DH5α /20LB(CP)
2/24 miniprep BclA-His-CBDcex1DH5α /20LB(CP)
2/24 miniprep BclA-His-CBDclos1DH5α /20LB(CP)
2/25 miniprep INP-His-CBDcex1DH5α /20LB(CP)
2/25 miniprep INP-His-CBDclos1DH5α /20LB(CP)

7/22

Transformation

 Yamamoto
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
2/4 parts awakening E00401DH5α /20LB(AMP)

8/1

Electrophoresis

 Iguchi
Lanesample/(&microl)Length(bp)
18/1 colonyPCR BclA control colony4 /10672
28/1 colonyPCR BclA control colony5 /10672
38/1 colonyPCR BclA control colony6 /10672
48/1 colonyPCR INP control colony4 /101563
58/1 colonyPCR INP control colony5 /101563
68/1 colonyPCR INP control colony6 /10
71kb ladder /5

8/1

Restriction Enzyme Digestion

 Iguchi
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
8/1 miniprep BclA-scFv8.36010110×H 3313.6430
8/1 miniprep BclA-CBDcex colony118.2001110×H /306.830
8/1 miniprep BclA-CBDclos colony121.3001110×M /303.730
8/1 miniprep INP-CBDcex colony118.0001110×M /307.030
8/1 miniprep INP-CBDclos colony110.8001110×M /3016.2

8/2

Gel Extraction

 Yamamoto
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
1BclA-scFv XPcut30
2BclA-CBDcex SPcut30
3BclA-CBDclos SPcut30
4INP-CBDcex SPcut30
5INP-CBDclos SPcut30
61kb ladder 30

8/2

Gel Extraction

 Yamamoto
Sample Concentration ng/&microl 260/280 260/230
8/2 Electrophoresis BclA-scFv XPcut 1.0 23.20 0.12
8/2 Electrophoresis BclA-CBDcex SPcut 10.8 1.89 1.35
8/2 Electrophoresis BclA-CBDclos SPcut 18.0 1.92 1.88
8/2 Electrophoresis INP-CBDcex SPcut 25.4 1.93 1.81
8/2 Electrophoresis INP-CBDclos SPcut 60.9

8/2

Ligation

 Yamamoto
Vector/(&microl)Insert/(&microl)Ligation High/(&microl)Total/(&microl)
8/2 purification BclA-CBDcex SPcut /3.58/2 purification BclA-scFv XPcut /47.515
8/2 purification BclA-CBDclos SPcut /3.58/2 purification BclA-scFv XPcut /47.515
8/2 purification INP-CBDcex SPcut /3.58/2 purification BclA-scFv XPcut /47.515
8/2 purification INP-CBDclos SPcut /3.58/2 purification BclA-scFv XPcut /47.5

8/2

Transformation

 Yamamoto, Yamada
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
8/2 ligation BclA-scFv+BclA-CBDcex2DH5α /20LB(CP)
8/2 ligation BclA-scFv+BclA-CBDclos2DH5α /20LB(CP)
8/2 ligation BclA-scFv+INP-CBDcex2DH5α /20LB(CP)
8/2 ligation BclA-scFv+INP-CBDclos2DH5α /20

8/2

Liquid Culture

 Yamada
Namemedium
BclA-CBDcex glycerol stockLB(CP)
BclA-CBDclos glycerol stockLB(CP)
INP-CBDcex glycerol stockLB(CP)
INP-CBDclos glycerol stock

8/3

Liquid Culture

 Yamada
Namemedium
7/28 transformation BclA control colony5LB(CP)
7/28 transformation BclA control colony6LB(CP)
7/28 transformation INP control colony4LB(CP)
7/28 transformation INP control colony5
7/28 transformation INP control colony6

8/3

Restriction Enzyme Digestion

 Yamamoto
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
8/1 miniprep BclA-scFv8.36010110×H /3313.6430
8/1 miniprep BclA-CBDcex18.2001110×H /306.830
8/1 miniprep BclA-CBDclos21.3001110×M /303.730
8/1 miniprep INP-CBDcex 18.0001110×M /307.030
8/1 miniprep INP-CBDclos10.8001110×M /3016.2

8/3

Ligation

 Yamamoto
Vector/(&microl)Insert/(&microl)Ligation High/(&microl)Total/(&microl)
8/1 Restriction Enzyme Digestion BclA-CBDcex /3.58/1 Restriction Enzyme Digestion BclA-scFv /47.515
8/1 Restriction Enzyme Digestion BclA-CBDclos /3.58/1 Restriction Enzyme Digestion BclA-scFv /47.515
8/1 Restriction Enzyme Digestion INP-CBDcex /3.58/1 Restriction Enzyme Digestion BclA-scFv /47.515
8/1 Restriction Enzyme Digestion INP-CBDclos /3.58/1 Restriction Enzyme Digestion BclA-scFv /47.515

8/3

Transformation

 Yamada
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
8/3 ligation BclA-scFv+BclA-CBDcex15DH5α /20LB(CP)
8/3 ligation BclA-scFv+BclA-CBDclos15DH5α /20LB(CP)
8/3 ligation BclA-scFv+INP-CBDcex15DH5α /20LB(CP)
8/3 ligation BclA-scFv+INP-CBDclos15DH5α /20

8/4

Colony PCR (typeA)

 Yamamoto
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
8/3 transformation BclA-scFv+BclA-CBDcex colony1VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+BclA-CBDcex colony2VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+BclA-CBDcex colony3VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+BclA-CBDcex colony4VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+BclA-CBDclos colony1VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+BclA-CBDclos colony2VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+BclA-CBDclos colony3VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+BclA-CBDclos colony4VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+INP-CBDcex colony1VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+INP-CBDcex colony2VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+INP-CBDcex colony3VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+INP-CBDcex colony4VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+INP-CBDclos colony1VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+INP-CBDclos colony2VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+INP-CBDclos colony3VF 1.25 VR1.251012.525
8/3 transformation BclA-scFv+INP-CBDclos colony4VF 1.25 VR1.251012.525

8/4

PCR (Steps)

 Yamamoto
StepsTemparatureTimeCycle
Predenature95℃1min1
Denature95℃15sec30
Annealing60℃15sec30
Extension72℃10sec30
Final Elongation72℃5min1
Final Holding4℃--

8/4

Miniprep

 Yamada
DNAConcentration/(&microg/ml)260/280260/230
8/3 liquid culture BclA control colony5137.21.771.03
8/3 liquid culture BclA control colony6117.61.871.54
8/3 liquid culture INP control4165.21.871.68
8/3 liquid culture INP control5150.61.901.81
8/3 liquid culture INP control6147.41.79

8/4

Sequence

 Yamamoto, Yamada
No primer(10&microM) MilliQ Total
1 / 2 BclA_5_VF(2.18&microl) / BclA_5_VR(2.18&microl) 2&microl / 2&microl 5.82&microl / 5.82&microl 10&microl
3 / 4 BclA_6_VF(2.55&microl) / BclA_6_VR(2.55&microl) 2&microl / 2&microl 5.45&microl / 5.45&microl 10&microl
5 / 6 INP_4_VF(1.81&microl) / INP_4_VR(1.81&microl) 2&microl / 2&microl 6.19&microl / 6.19&microl 10&microl
7 / 8 INP_5_VF(1.99&microl) / INP_5_VR(1.99&microl) 2&microl / 2&microl 6.01&microl / 6.01&microl 10&microl

8/4

Electrophoresis

 Yamada
Lanesample/(&microl)Length(bp)
18/4 colony PCR BclA-scFv+BclA-CBDcex colony1 /101716
28/4 colony PCR BclA-scFv+BclA-CBDcex colony2 /101716
38/4 colony PCR BclA-scFv+BclA-CBDcex colony3 /101716
48/4 colony PCR BclA-scFv+BclA-CBDcex colony4 /101716
58/4 colony PCR BclA-scFv+BclA-CBDclos colony1 /101716
68/4 colony PCR BclA-scFv+BclA-CBDclos colony2 /101716
78/4 colony PCR BclA-scFv+BclA-CBDclos colony3 /101716
88/4 colony PCR BclA-scFv+BclA-CBDclos colony4 /101716
9100bp ladder /5-
101kb ladder /5-
11--
12--

8/4

Electrophoresis

 Yamada
Lanesample/(&microl)Length(bp)
18/4 colony PCR BclA-scFv+INP-CBDcex colony1 /102606
28/4 colony PCR BclA-scFv+INP-CBDcex colony2 /102606
38/4 colony PCR BclA-scFv+INP-CBDcex colony3 /102606
48/4 colony PCR BclA-scFv+INP-CBDcex colony4 /102606
58/4 colony PCR BclA-scFv+INP-CBDclos colony1 /102606
68/4 colony PCR BclA-scFv+INP-CBDclos colony2 /102606
78/4 colony PCR BclA-scFv+INP-CBDclos colony3 /102606
88/4 colony PCR BclA-scFv+INP-CBDclos colony4 /102606
9100bp ladder /5-
101kb ladder /5
11--
12--

8/5

PartsAwaking

 Yamamoto
DescriptionPartNameBackboneWellPlateYearPlateNumberMilliQ(&microl)
RFPJ23105J6100217N20164

8/5

Transformation

 Yamamoto
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
8/5 Parts awakening J231051&microlDH5α /20

8/9

Restriction Enzyme Digestion

 Iguchi, Yamada
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
8/1 miniprep BclA-scFv8.36010110×H /3313.6930
8/1 and 7/25 miniprep BclA-CBDcex18.001110×H /306.830
2/25 miniprep BclA-CBDclos21.3001110×M /303.730
2/25 and 7/25 minniprep INP-CBDcex18.0001110×M /307.030
8/1 miniprep INP-CBDclos10.8001110×M /3016.230

8/9

Liquid Culture

 Yamada
Namemedium
8/1 transformation J23105

8/9

Miniprep

 Yamada
DNAConcentration/(&microg/ml)260/280260/230
8/3 liquid culture BclA-CBDccex225.81.741.63
8/3 liquid culture BclA-CBDclos102.31.801.66
8/3 liquid culture INP-CBDcex112.41.861.77
8/3 liquid culture INP-CBDclos42.81.760.78

8/9

Electrophoresis

 Yamada
Lanesample/(&microl)Length(bp)

8/10

Gel Extraction

 Yamada
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
A11kb ladder10
A2BclA-scFv 8/9XPcut30
A3BclA-CBDcex 8/9 SPcut30
A4BclA-CBDclos 8/9 SPcu30
---
B11kb ladder10
B2INP-CBDcex 8/9 SPcut30
B3INP-CBDclos 8/9 SPcut

8/10

Gel Extraction(Measurement)

 Yamada
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
BclA-scFv 8/9XPcut2.61.130.58
BclA-CBDcex 8/9 SPcut6.31.540.97
BclA-CBDclos 8/9 SPcut4.71.490.48
INP-CBDcex 8/9 SPcut4.41.390.67
INP-CBDclos 8/9 SPcut3.51.29

8/10

Miniprep

 Notsu
DNAConcentration/(&microg/ml)260/280260/230
8/9 liquid culture J23105 (AMP) RFP colony3178.81.981.92

8/12

Liquid Culture

 Iguchi
Namemedium
8/2 liquid culture INP-CBDclosLB(CP)
BclA-scFv glycerol stock 1LB(CP)
BclA-scFv glycerol stock 2LB(CP)
BclA-scFv glycerol stock 3LB(CP)
BclA-scFv glycerol stock 4LB(CP)

8/18

Miniprep

 Notsu
DNAConcentration/(&microg/ml)260/280260/230
8/12 liquid culture INP-CBDclos45.31.821.31
8/12 liquid culture BclA-scFv glycerol stock 1101.61.590.71
8/12 liquid culture BclA-scFv glycerol stock 2100.71.670.78
8/12 liquid culture BclA-scFv glycerol stock 377.81.600.70
8/12 liquid culture BclA-scFv glycerol stock 441.01.871.27

8/20

Liquid Culture

 Ohara, Isoda
Namemedium
BclA-CBDcexLB(CP)
BclA-CBDclosLB(CP)
INP-CBDcexLB(CP)
INP-CBDclos

8/21

Liquid Culture

 Ohara
Namemedium
BclA-scFv ①(glisto）LB(CP)
BclA-scFv ②LB(CP)
BclA-scFv ③LB(CP)
BclA-scFv ④

8/22

Electrophoresis

 Sassa
Lanesample/(&microl)Length(bp)
8/22 Restraction Ensyme Digestion 1.1kb ladder5
8/22 Restraction Ensyme Digestion Bcla-cex252520
8/22 Restraction Ensyme Digestion Bcla-clos252520
8/22 Restraction Ensyme Digestion INP-cex(s,pcut)253500
8/22 Restraction Ensyme Digestion INP-clos253500
8/22 Restraction Ensyme Digestion INP-cex252541
8/22 Restraction Ensyme Digestion J23105 (AMP) RFP 8/21 old25950
8/22 Restraction Ensyme Digestion Liquid culture Bcla-scfv glisto 425932
8/22 Restraction Ensyme Digestion Bcla-scfv glisto 225932
100ladder5

8/22

Restriction Enzyme Digestion

 Ohara
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
8/12 Liquid cultureBclA-scFv glisto ④3801015550
8/12 Liquid cultureBclA-scFv glisto ②38010155050

8/22

Liquid Culture

 Yoshimoto, Ohara, Isoda
Namemedium
BclA controlLB(CP)
INP control

8/22

Transformation

 Ohara
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
2/9 J23105 RFPDH5α /20

8/22

CBD

 Yoshimoto
1st
samplebeforeafterwash
BclAclos+0.208/0.206/0.2060.131/0.135/0.140.111/0.112/0.109
INPclos+0.183/0.186/0.1840.239/0.23/0.220.158/0.155/0.169

8/22

CBD

 Yoshimoto
2nd
samplebeforeafterwash
BclAclos+0.208/0.205/0.2040.078/0.074/0.0750.056/0.056/0.039
INPclos+0.183/0.181/0.1870.093/0.097/0.0990.067/0.073/0.069

8/23

CBD

 Yamada
2nd
samplebeforeafterwash
BclAcex+ (15min)0.247/0.247/0.2480.221/0.224/0.2180.085/0.075/0.073
BclAcex+ (30min)0.247/0.247/0.2480.243/0.229/0.2280.034/0.048/0.038
BclAcex+ (60min)0.247/0.247/0.2480.224/0.221/0.2170.065/0.064/0.065
BclAcex- (15min)0.247/0.247/0.2480.257/0.253/0.258
BclAcex- (30min)0.247/0.247/0.2480.236/0.230/0.235
BclAcex- (60min)0.247/0.247/0.2480.249/0.248/0.247
cellulose only (60min)0.021/0.021/0.023

8/23

CBD

 Yamada
4th
samplebeforeafterwash
BclAcex+ (2ml tube)0.227/0.226/0.2250.108/0.108/0.1080.076/0.074/0.081
BclActl+ (2ml tube)0.186/0.203/0.1890.082/0.081/0.0820.055/0.046/0.044
BclAcex- (2ml tube)0.235/0.237/0.2330.261/0.262/0.254
BclAcex+ (1.5ml tube)0.237/0.226/0.2360.159/0.157/0.1580.066/0.063/0.065
BclActl+ (1.5ml tube)0.177/0.173/0.1790.112/0.109/0.1130.080/0.076/0.095
BclAcex- (1.5ml tube)0.240/0.235/0.2380.255/0.254/0.253

8/24

Liquid Culture

 Sassa
Namemedium
8/5 transormation J2305

8/24

Liquid Culture

 Ohara
Namemedium
8/24 BclA-scFv 1LB(CP)
8/24 BclA-scFv 2LB(CP)
8/24 BclA-scFv 3LB(CP)
8/24 BclA-scFv 4

8/29

Miniprep

 Sassa
DNAConcentration/(&microg/ml)260/280260/230
1. 8/23Liquidculture Bcla scFv111.61.981.82
2. 8/23Liquidculture Bcla scFv 143.12.002.03
2'. 8/23Liquidculture Bcla scFv141.62.042.10
3. 8/23Liquidculture Bcla scFv145.22.072.01
8/23Liquidculture J2310563.12.011.69
8/23Liquidculture J2310568.12.00

8/24

CBD

 Isoda
5th
samplebeforeafterwash
INPcex+0.195/0.191/0.1920.103/0.102/0.1090.059/0.051/0.055
INPcex-0.190/0.191/0.2050.186/0.187/0.186
INPclos+0.187/0.187/0.1880.121/0.122/0.1260.051/0.058/0.059
INPclos-0.196/0.195/0.1910.206/0.204/0.202
INPctl+0.172/0.163/0.1620.116/0.177/0.1240.066/0.063/0.062
INPctl-0.171/0.175/0.1650.167/0.182/0.171

8/24

CBD

 Isoda
6thnullnullnull
samplebeforeafterwash
INPcex+0.184/0.184/0.1890.104/0.106/0.1060.044/0.044/0.056
INPcex-0.186/0.190/0.1840.182/0.181/0.184
INPclos+0.184/0.189/0.1850.097/0.098/0.0990.037/0.037/0.038
INPclos-0.187/0.192/0.1900.185/0.184/0.189
INPctl+0.173/0.184/0.1800.095/0.099/0.0950.045/0.046/0.046
INPctl-0.185/0.184/0.1830.175/0.175/0.175
BclAcex+0.170/0.168/0.1740.099/0.094/0.0920.037/0.032/0.036
BclAcex-0.186/0.189/0.1710.164/0.167/0.164
cellulose only0.006/0.005/0.0060.013/0.010/0.006

8/25

CBD

 Yamada
7thnullnullnull
samplebeforeafterwash
BclAcex+0.181/0.181/0.1810.192/0.196/0.1950/0/0
BclAcex-0.149/0.150/0.1510.221/0.213/0.211
BclActl+0.169/0.165/0.1670.154/0.156/0.1570.012/0.013/0.011
BclActl-0.151/0.153/0.1540.187/0.178/0.173

8/26

PCR (Target)

 Sassa
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
8/10 miniprep J23105 /3VF /1.25VR /1.25-7KAPA2GFAST /12.525
8/24 OD0.206 INP-CBDclos /3VF /1.25VR /1.25-7KAPA2GFAST /12.525
8/24 OD0.202 INP-CBDcexVF /1.25VR /1.25-7KAPA2GFAST /12.525

8/26

Electrophoresis

 Yamamoto, Sassa
Lanesample/(&microl)Length(bp)
11kb ladder /5
28/26 PCR INP-CBDcex VFVR /15
38/26 PCR INP-CBDclos VFVR /15

8/26

Miniprep

 Sassa
DNAConcentration/(&microg/ml)260/280260/230
8/26 Gel Extraction Bcla cex8.31.520.25
8/26 Gel Extraction Bcla clos10.21.700.26
8/26 Gel Extraction INP cex11.41.760.15
8/26 Gel Extraction INP clos7.71.620.09
8/26 Gel Extraction Bcla scFv glisto7.01.590.20
8/26 Gel Extraction Bcla scFv glisto14.16.33

8/26

Miniprep

 Yamamoto, Sassa
DNAConcentration/(&microg/ml)260/280260/230
8/26 Gel Extraction Bcla cex8.31.520.25
8/26 Gel Extraction Bcla clos10.21.700.26
8/26 Gel Extraction INP cex11.41.760.15
8/26 Gel Extraction INP clos7.71.620.09
8/26 Gel Extraction Bcla scFv glisto7.01.590.20
8/26 Gel Extraction Bcla scFv glisto14.16.33

8/26

CBD

 Yoshimoto
8thnullnullnull
samplebeforeafterwash
BclAcex+0.183/0.185/0.1880.116/0.115/0.1140.086/0.086/0.089
BclActl+0.116/0.115/0.1140.096/0.092/0.0870.09/0.079/0.084
cellulose only0.037/0.035/0.037

8/26

CBD

 Ohara
9thnullnullnull
samplebeforeafterwash
BclAcex+0.186/0.186/0.1840.170/0.169/0.1700.021/0.024/0.020
BclAcex-0.154/0.149/0.1490.142/0.143/0.141
BclActl+0.194/0.189/0.1890.167/0.164/0.1590.010/0.009/0.008
BclActl-0.162/0.168/0.1690.159/0.162/0.165

8/29

Ligation

 Sassa
Insert DNAvector DNALigation HighTotal
8/26 Gel Extraction BclAscFv /3.48/26 Gel Extraction Bcla CBDcex /4.17.515
8/26 Gel Extraction BclAscFv /3.18/26 Gel Extraction Bcla CBD clos /4.47.515
8/26 Gel Extraction BclAscFvBclAscFv /3.88/26 Gel Extraction Bcla CBD cex /4.77.515
8/26 Gel Extraction BclAscFv /2.68/26 Gel Extraction Bcla CBD clos /4.97.5

8/29

Western Blotting

 Ohara, Sassa
lanesample
1Marker
2Sano VLP 0.5&microl
3Sano VLP 0.2&microl
4Sano VLP 0.1&microl
5Sano VLP 0.05&microl
6Sano VLP 0.02&microl
7Sano VLP 0.05&microl

blotting CC
1'antibody time/buffer 1:3000 6ml 1h
2'antibody time/buffer 1:10000 10ml 1h
Blocking 1day
transfer 100mA 1h

8/29

PCR (Target)

 Miyazaki
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
BclA-CBDcexVF /1.25VR /1.25-10KAPA2GFAST /12.525
BclA-CBDclosVF /1.25VR /1.25-10KAPA2GFAST /12.525
INP-CBDcexVF /1.25VR /1.25-10KAPA2GFAST /12.525
INP-CBDclosVF /1.25VR /1.25-10KAPA2GFAST /12.525

8/29

ligation

 Miyazaki
Insert DNAVector DNALigation High Total
BclAscFv XPcut 6.3INPCBDcex SPcut 1.27.515
BclAscFv XPcut 6.4INPCBDclos SPcut 1.17.515

8/29

Miniprep

 Sassa, Miyazaki
DNAConcentration/(&microg/ml)260/280260/230
BclA scFv 18.41.630.11
BclA scFv 25.61.910.04
INP CBDcex44.81.900.70
INP CBDclos33.11.820.68
BclA cex26.31.930.62
BclA clos 24.41.930.66
J23105 VF VR33.21.880.24

8/1

Colony PCR (typeA)

 Yamamoto
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
7/28 transformaton BclA control colony4VF/1.25 VR/1.251012.525
7/28 transformaton BclA control colony5VF/1.25 VR/1.251012.525
7/28 transformaton BclA control colony6VF/1.25 VR/1.251012.525
7/28 transformaton INP control colony4VF/1.25 VR/1.251012.525
7/28 transformaton INP control colony5VF/1.25 VR/1.251012.525
7/28 transformaton INP control colony6VF/1.25 VR/1.251012.5

8/1

PCR (Steps)

 Yamamoto
StepsTemparatureTimeCycle
Predenature95℃1min1
Denature95℃15sec30
Annealing60℃15sec30
Extension72℃10sec30
Final Elogation72℃5min1
Final Holding-1

8/1

Miniprep

 Yamamoto
DNAConcentration/(&microg/ml)260/280260/230
7/28 liquid culture BclA-scFv239.11.871.78
7/28 liquid culture INP-scFv colony1263.41.861.99
7/28 liquid culture E0040 colony198.81.842.15
7/28 liquid culture BclA-CBDcex colony1109.91.821.37
7/28 liquid culture BclA-CBDclos colony194.11.891.93
7/28 liquid culture INP-CBDcex colony1111.41.821.39
7/28 liquid culture INP-CBDclos colony1185.11.81

9/5

Liquid Culture

 Isoda
Namemedium
BclA-CBDcex /20mlLB(CP)
BclA-control /20mlLB(CP)

9/5

Gel Extraction

 Sassa, Miyazaki
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
9/5 electrophoresis 2 J23105 RFP XPcut backbone
9/5 electrophoresis 2J23105 RFP XPcut insert
9/5 electrophoresis 1BclAcex SPcut
9/5 electrophoresis 2BclAclos SPcut
9/5 electrophoresis 3INPcex SPcut
9/5 electrophoresis 4INPclos SPcut

9/6

Colony PCR (typeA)

 Yamamoto
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
Bclascfv+BclaCBDcex colony1vf1.25 vr1.2510KAPA2GFAST 12.525
〃colony2〃〃〃〃
〃colony3〃〃〃〃
〃colony4〃〃〃〃
Bclascfv+BclaCBDclos colony1〃〃〃〃
〃colony2〃〃〃〃
〃colony3〃〃〃〃
〃colony4〃〃〃〃
Bclascfv+INPCBDcex colony1〃〃〃〃
〃colony2〃〃〃〃
〃colony3〃〃〃〃
〃colony4〃〃〃〃
Bclascfv+INPCBDclos colony1〃〃〃〃
〃colony2〃〃〃〃
〃colony3〃〃〃onaji
〃colony4〃〃〃〃

9/6

Ligation

 Yamamoto
Insert DNA(&microl)Vector DNA/(&microl)Ligation High/(&microl)Total/(&microl)
9/5 Gel Extraction BclA cex SPcut/79/5 Gel Extraction J23105 xpcut 36/0.57.515
9/5 Gel Extraction BclA clos SPcut/79/5 Gel Extraction J23105 xpcut 36/0.57.515
9/5 Gel Extraction INP cex SPcut/79/5 Gel Extraction J23105 xpcut 36/0.57.515
9/5 Gel Extraction INP clos SPcut/79/5 Gel Extraction J23105 xpcut 36/0.57.515

9/6

Electrophoresis
Lanesample/(&microl)Length(bp)
11kb ladder/50
29/6 PCR scfv+Bclacex colony1/150
3〃2/150
4〃3/150
5〃4/150
69/6 PCR scfv+Bclaclos colony10
7〃20
8〃30
9〃40
109/6 PCR scfv+INPcex colony10
11〃20
12〃30
13〃40
149/6 PCR scfv+INPclos colony10
15〃20
16〃30
17〃4

9/6

Western Blotting

 Ohara, Sassa
lanesample
1Marker
2Sano VLP 0.5&microl
3Sano VLP 0.2&microl
4Sano VLP 0.1&microl
5Sano VLP 0.05&microl
6Sano VLP 0.02&microl
7Sano VLP 0.05&microl

blotting CC
1'antibody time/buffer 1:3000 6ml 1h
2'antibody time/buffer 1:10000 10ml 1h
Blocking 1day
transfer 100mA 1h

9/6

CBD

 Isoda
10thnullnullnull
samplebeforeafterwash
BclAcex+0.2390.222
BclActl+0.2120.190
BclAcex-0.2390.233
BclActl-0.2210.210

9/6

CBD

 Isoda
11thnullnullnull
samplebeforeafterwash
BclAcex+ (NaCl)0.2020.176
BclAcex- (NaCl)0.2020.206
BclAcex- (NaCl)0.2020.264
BclAcex+ (PBS buffer)0.2120.179
BclAcex- (PBS buffer)0.2120.209
BclActl+ (PBS buffer)0.2110.172

9/7

Colony PCR (typeA)

 Yamamoto
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
Bclascfv-BclaCBDcex colony5vf12.5 vr12.510KAPA2GFASTHSRM 12.525
〃6====
〃7====
〃8====
〃9====
〃10====
9/5 Bclascfv-BclaCBDclos 5====
〃6====
9/5 Bclascfv-INPCBDcex1====
〃2====
〃3====
〃4====
Bclascfv-INPCBDclos1====
〃2====
〃3====
〃4===

9/7

Miniprep

 Yamamoto
DNAConcentration/(&microg/ml)260/280260/230
Bcla-CBDcex99.71.942.08
Bcla-CBDclos73.51.962.15
INP-CBDcex45.31.992.14
INP-CBDclos49.61.89

9/7

Restriction Enzyme Digestion

 Yamamoto
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
8/5 miniprep INP-CBDcex25001110×M/30030
8/5 miniprep INP-CBDclos25001110×M/30030
9/13 miniprep BclA-scFv122010110×H/33030

9/7

Liquid Culture

 Yamamoto
Namemedium
9/5 Transformation BclascFv-BclaCBDcex colony5LB(CP)/5ml
〃BclascFv-BclaCBDclos colony4

9/7

SEM

 Ohara
numbersampleOD600VLP(&microl)
1BclAscFv 3c50.9901
2BclAscFv 1c51.0011
3INPscFv 3c51.0621
4BclA CTL1.0531
5INP CTL0.9521

9/7

Liquid Culture

 Isoda
Namemedium
Bcla-CBDcexLB(CP)/30ml
Bcla-control

9/7

CBD

 Isoda
12thnullnullnull
samplebeforeafterwash
BclAcex+ (PBS buffer)0.195/0.1920.188/0.189
BclActl+ (PBS buffer)0.198/0.1950.192/0.189
BclAcex+ (CB buffer)0.186/0.1840.176/0.173
BclActl+ (CB buffer)0.172/0.1690.151/0.155

9/7

CBD

 Isoda
13thnullnullnull
samplebeforeafterwash
BclAcex+ (PBS buffer) (Bemliese)0.201/0.2010.201/0.201
BclActl+ (PBS buffer) (Bemliese)0.224/0.2230.218/0.215
BclAcex+ (CB buffer) (Bemliese)0.266/0.2570.243/0.242
BclActl+ (CB buffer) (Bemliese)0.178/0.1770.157/0.149
BclAcex+ (PBS buffer) (Kimwipe)0.203/0.2010.200/0.196
BclActl+ (PBS buffer) (Kimwipe)0.221/0.2200.224/0.229
BclAcex+ (CB buffer) (Kimwipe)0.273/0.2660.244/0.242
BclActl+ (CB buffer) (Kimwipe)0.172/0.1720.154/0.154

9/8

Gel Extraction

 Yamamoto
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder5
29/7 restriction enzyme design BclA scFv X/Pcut30
39/7 restriction enzyme design INP CBDcex S/Pcut30
49/7 restriction enzyme design INP CBDclos S/Pcut30
5100bp ladder5

9/8

Colony PCR (typeA)

 Yamamoto
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
9/5 TransformationINPcex-scFv colony5vf 1.25 vr1.2510KAPA2GFAST/12.525
〃colony6====
〃colony7====
〃colony8====
9/5 Transformation INPclos-BclascFv colony1====
〃colony2====
〃colony5====
〃colony6===

9/8

Western Blotting

 Ohara, Sassa
lanesample
1Marker
2Sano VLP 0.5&microl
3Sano VLP 0.2&microl
4Sano VLP 0.1&microl
5Sano VLP 0.05&microl
6Sano VLP 0.02&microl
7Sano VLP 0.05&microl

1'antibody time/buffer 1:3000 6ml 1h
2'antibody time/buffer 1:10000 10ml 1h
Blocking 1day
transfer 100mA 1h

9/8

Ligation

 Yamamoto
Insert/&microlVector/&microlLigation High/&microlTotal
9/8 Gel Extraction BclA scFv X/Pcut /69/8 Gel Extraction INP CBDcex S/Pcut /1.57.515
9/8 Gel Extraction BclA scFv X/Pcut /69/8 Gel Extraction INP CBDcex S/Pcut /1.57.515

9/8

Transformation

 Yamamoto, Sassa
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
9/8 Ligation Bclascfv-INPCBDcex2DH5a/20LB(CP)
〃BclascFv-INPCBDclos2〃〃
9/7Miniprep BclaCBDcex+8/24 Miniprep J231051〃LB(CP+AMP)
〃Bcla-CBDclos+8/24 J231051〃〃
9/7 Miniprep INP-CBDcex+8/24 Miniprep J231051〃〃
9/7 Miniprep INP-CBDclos+8/24 Miniprep J231051〃

9/8

Liquid Culture

 Isoda
Namemedium
Bcla-CBDcexLB(CP)/30ml
Bcla-CBDcontrol

9/8

CBD

 Isoda
14thnullnullnull
samplebeforeafterwash
CBDcex+ (CB buffer)0.241/0.2360.215/0.212
CBDctl+ (CB buffer)0.182/0.1800.159/0.159
BclAcex+ (CB buffer + Na2CO3)0.235/0.2350.177/0.177
BclActl+ (CB buffer+Na2CO3)0.227/0.2180.155/0.156

9/9

Glycerol Stock

 Yamamoto
sample /mlglycerol /ml
BclA control /0.80.2
BclA control /0.80.2

9/9

Glycerol Stock

 Yamamoto
sample /mlglycerol /ml
INP control /0.80.2
INP control /0.80.2

9/9

Liquid Culture

 Yamamoto
Namemedium

9/9

Colony PCR (typeA)

 Iguchi
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
INPcex-scFv colony9vf1.25 vr1.258.5KAPA2GFAST 12.525
〃colony10〃〃〃〃
〃colony11〃〃〃〃
9/5 transformation INPclos-scFv colony1〃〃〃〃
〃colony5〃〃〃

9/9

Liquid Culture

 Miyazaki
Namemedium
scFv lc3LB()

9/9

Western Blotting

 Ohara, Sassa
lanesample
1Marker
2Sano VLP 0.5&microl
3Sano VLP 1&microl
4Sano VLP 0.2&microl
5Sano VLP 0.1&microl
6Sano VLP 0.05&microl
7Sano VLP 0.02&microl
8Sano VLP 0.5&microl 95℃ without boiling
9only SDS lysis buffer

1'antibody time/buffer 1:3000 9ml 1h
2'antibody time/buffer 1:10000 10ml 1h
Blocking 1h
transfer 100mA 1h

9/9

Miniprep

 Miyazaki
DNAConcentration/(&microg/ml)260/280260/230
scFv Bcla/1ul271.82.092.15
VLP/1ul1.671.98

9/9

Colony PCR (typeA)

 Yamamoto
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
BclaCBDcexJ23105vf1.25 vr1.2510KAPA2GFAST HSRM withDye/12.525
9/6 Transformation BclaCBDclos J23105〃〃〃〃
〃INPCBDcex J23105〃〃〃〃
〃INPCBDcexJ23105〃〃〃〃

9/9

Transformation
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
2/24 miniprep Bcla-his-cex colony20.1DH5a/20ul

9/9

Liquid Culture

 Iguchi
Namemedium
9/5 transformation INPcex-BclascFv colony9LB(CP)/5ml
〃INPclos-BclascFv colony1

9/9

Colony PCR (typeA)

 Yamamoto
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
9/5 transformation Bcla CBDcex J23105 vf1.0 vr1.023Quick Taq/2550
〃BclaCBDclos J23105〃〃〃〃
〃INPCBDcex J23105〃〃〃〃
〃INPCBDclos J23105〃〃〃

9/12

Restriction Enzyme Digestion

 Miyazaki
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
9/9miniprep scFv Bcla lc3 29ul10110×H/559

9/12

Electrophoresis

 Miyazaki
Lanesample/(&microl)Length(bp)
11kb ladder/5 -
2Bcla CBDcex j23105/15①789 1140
3〃②〃
4Bcla CBDclos J23105①757 1140
5〃②〃
6INP CBDcex J23105①1664 1140
7〃②〃
8INP CBDclos J23105①1796 1140
9〃②〃
10100bp ladder

9/12

Colony PCR (typeA)

 Miyazaki
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
Bcla CBDcex J23105①vf0.5 vr0.511.5Quick Taq/12.525
〃②〃〃〃〃
〃③〃〃〃〃
〃④〃〃〃〃
Bcla CBDclos J23105①〃〃〃〃
〃②〃〃〃〃
〃③〃〃〃〃
〃④〃〃〃

9/12

Colony PCR (typeA)

 Miyazaki
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
INP CBDcex J23105vf0.5 vr0.511.5Quick Taq12.525
INP〃②====
〃③====
〃④====
〃clos①====
〃②====
〃③====
〃④===

9/13

Western Blotting

 Ohara, Sassa
lanesample
1Marker
2Sano VLP 1&microl
3Sano VLP 1&microl
4Sano VLP 0.2&microl
5Sano VLP 0.1&microl
6Sano VLP 0.05&microl
7Sano VLP 0.02&microl
8Sano VLP 0.5&microl 95℃ without boiling
9only SDS lysis buffer

1'antibody time/buffer 1:3000 9ml 1h
2'antibody time/buffer 1:10000 10ml 1h
Blocking 1h
transfer 100mA 1h
30ml TBS-T/blocking reagent 1.5g

9/14

Liquid Culture

 Isoda
Namemedium
9/9 transformation BclA-CBDcexLB(CP)
8/24 transformation BclA-CBDclosLB(CP)
8/24 transformation INP-CBDcexLB(CP)
8/20 transformation LB(CP)

9/14

Western Blotting

 Ohara, Sassa
lanesample
1Marker
2Sano VLP 0.5&microl＋INP-scFv 100&microl
3Sano VLP 0.5&microl＋BAN-scFv 100&microl
4Sano VLP 0.2&microl ＋BAN-scFv 100&microl
5Sano VLP 0.2&microl ＋BAN-scFv 70&microl ＋BANcontrol30&microl
6Sano VLP 0.2&microl ＋BAN-scFv 20&microl ＋BANcontrol80&microl
7Sano VLP 0.2&microl ＋BANcontrol100&microl
8Sano VLP 0.2&microl 95℃ without boiling
9Sano VLP 0.5&microl 95℃ without boiling
10Marker

1'antibody time/buffer 1:1000 10ml / Blocking buffer　overnight
2'antibody time/buffer 1:2500 10ml / Blocking buffer 1h
Blocking 1h
transfer 100mA 1h
25ml TBS-T/blocking reagent 1.3g

9/15

Western Blotting

 Ohara, Sassa
lanesample
1Marker
2Sano VLP 0.5&microl＋INP-scFv 100&microl
3Sano VLP 0.5&microl＋BAN-scFv 100&microl
4Sano VLP 0.2&microl ＋BAN-scFv 100&microl
5Sano VLP 0.2&microl ＋BAN-scFv 70&microl ＋BANcontrol30&microl
6Sano VLP 0.2&microl ＋BAN-scFv 20&microl ＋BANcontrol80&microl
7Sano VLP 0.2&microl ＋BANcontrol100&microl
8Sano VLP 0.2&microl 95℃ without boiling
9Sano VLP 0.5&microl 95℃ without boiling
10Marker

1'antibody time/buffer 1:1000 10ml / Blocking buffer　overnight
2'antibody time/buffer 1:2500 10ml / Blocking buffer 1h
Blocking 1h
transfer 100mA 1h
25ml TBS-T/blocking reagent 1.3g

9/15

Liquid Culture

 Sassa
Namemedium
9/7 BclA scFv 3C5 9/7 2 LB(CP) /20mL
9/14 BclA cex (9/9 tranceformation)LB(CP) /20mL
9/7 INP scFv 3C5LB(CP) /20mL
9/9 BclA scFv 1c3LB(CP) /20mL
9/7 INP CTLLB(CP) /20mL
9/7 BclA CTL 2LB(CP) /20mL

9/15

Liquid Culture

 Ohara, Isoda
Namemedium
BclA CBDclos glycerol stockLB(CP)
INP-CBDcex glycerol stockLB(CP)
INP-CBDclos glycerol stockLB(CP)

9/16

Transformation

 Uchino, Sassa
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
8/20 J23105 RFP 1/100 20DH5α /20LB(CP)

9/19

Transformation

 Sassa, Takimoto
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
2/24 BAN-cex1DH5α /20LB(CP)
8/5 INP-cex1DH5α /20LB(CP)
2/8 INP HIsR1DH5α /20LB(CP)
2/19 BAN-clos1DH5α /20LB(CP)
8/5 INP-clos1DH5α /20LB(CP)
2/4 E10101DH5α /20
8/18 BclA-scFv1DH5α /20LB(CP)

9/19

Liquid Culture

 Sassa, Takimoto
Namemedium
9/15 Liquid culture BclA scFv 3C5②LB(CP) /20
9/19 Liquid culture　Bcla CTLLB(CP) /20
9/15 Liquid culture INP-scFv 3C5LB(CP) /20
9/15 Liquid culture INP CTL

9/20

Liquid Culture

 Sassa
Namemedium
9/19 liquid culture BclA-scFv 3C5-2LB(CP)
9/19 liquid culture BclA-ctlLB(CP)
9/19 liquid culture INP-scFv 3C5LB(CP)
9/19 liquid culture INP-ctlLB(CP)

9/20

Western Blotting

 Ohara, Sassa
lanesample
MarkerM.Marker
1INPNC-6His-scFv 300 &microl＋VLP3 &microl
2INPNC-6His-scFv 300 &microl＋PBS3 &microl
3INPNC-6His-ctl300 &microl＋VLP3 &microl
4BclA-6His-scFv 300 &microl＋VLP3 &microl
5BclA-6His-scFv 300 &microl＋PBS3 &microl
6BclA-6His-ctl 300 &microl＋VLP3 &microl
7VLP0.5 &microl
8VLP0.2 &microl
9

Transfer ;60 min 100 mA(CC)
Blocking ;1h
1’antibody ;anti-NoV pAb(mouse)
1:1000 with blocking buffer O/N
2’antibody ;anti-mouse HRP　
1:2500 with blocking buffer 1 h
Detection time　80 sec
(ECL Prime)

9/20

Transformation

 Uchino
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
BclA-clos glycerol stock-DH5α /20LB(CP)
INP-clos glycerol stock-DH5α /20LB(CP)

9/21

PCR (Steps)

 Uchino
StepsTemparatureTimeCycle
Predenature95℃5min1
Denature98℃20sec30
Annealing65℃15sec30
Extension72℃30sec30
Final Elongation72℃3min1
Final Holding8℃--

9/21

PCR (Target)

 Uchino
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/8 miniprep INP-His9 /0.2VF /0.75m.scFv Rv /0.75-10.8KAPA HiFi Ready Mix /12.525
2/8 miniprep INP-His9 /0.2m.scFv Fw /0.75VR /0.75-10.8KAPA HiFi Ready Mix /12.525

VF 10&microM VR 10&microM

9/21

Gel Extraction

 Uchino
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
11kb ladder5
2100bp ladder 5
39/21 PCR INP-His9-125
49/21 PCR INP-His9-225

9/21

Liquid Culture

 Ohara, Sassa
Namemedium
9/19 Liquid culture BAN-scFv 3C5②LB(CP)
9/19 Liquid culture BAN-CTL LB(CP)
9/19 Liquid culture INP-scFv 3C5LB(CP)
9/19 Liquid culture INP-CTLLB(CP)
9/15 BAN- scFv 1C3LB(CP)
9/15 BAN-CBDcexLB(CP)

9/21

Western Blotting

 Ohara, Sassa
lanesample
MarkerM.Marker
1INPNC-6His-scFv 200 &microl＋VLP2 &microl
2INPNC-6His-scFv 200 &microl＋PBS2&microl(NC)
3INPNC-6His-ctl200 &microl＋VLP2 &microl(NC)
4BclA-6His-scFv200 ul(Vector pSB3C5)＋VLP2 ul
5BclA-6His-scFv 200 &microl＋VLP2 &microl (NC)
6 BclA-6His-scFv(Vector pSB1C3)＋VLP2 ul
7 BclA-6His-CBDcex 200 ul ＋ VLP2 ul (NC)
8PBS200 ul ＋VLP2 ul (NC)
9VLP1 ul (PC)

Transfer ;60 min 100 mA(CC)
Blocking ;1h
1’antibody ;anti-NoV pAb(mouse)
1:1000 with blocking buffer O/N
2’antibody ;anti-mouse HRP　
1:2500 with blocking buffer 1 h
Detection time　60 sec
(ECL Prime)

9/21

Gel Extraction(Measurement)

 Uchino
Sample/(&microl)Concentration(&microg/&microl)260/280260/230
9/21 PCR INP-His9-119.31.630.54
9/21 PCR INP-His9-248.61.820.24

9/21

PCR (Steps)

 Uchino
StepsTemparatureTimeCycle
Predenature95℃5min1
Denature98℃20sec30
Annealing65℃15sec30
Extension72℃30sec30
Final Elongation72℃3min1
Final Holding8℃-

9/21

PCR (Target)

 Uchino
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
9/21 Gel Extract INP-His9-1 /1&microl+9/21 Gel Extract INP-His9-2 /0.5&microlVF /0.75 VR /0.75-9.5KAPA HiFi /12.525

9/22

growth carve

 Sassa
sample0h0.5h1h1.5h2h3h4h5h6h7h8h
BAM scFv　①-0.048-0.047-0.0481.245-0.317-0.308-0.149
BAM scFv　②-0.0361.512-0.830-0.309-0.314-0.127
INP scFv　①-0.047-0.050-0.6441.045-0.307-0.153-0.142
INP scFv　②-0.038-0.o25-0.186-0.2800.000-0.289
BAN CBDcex　①-0.045-0.048-0.0441.089-0.322-0.284-0.296
BAN CBDcex　②-0.0341.204-1.057-0.3110.268-0.285
BAN CBDclos　①0.035-0.037-0.0351.261-0.323-0.160-0.287
BAN CBDclos　②-0.0181.306-0.050-0.282-0.128-0.293
INP CBDcex　①-0.020-0.027-0.0161.245-0.295-0.263-0.288
INP CBDcex　②-0.0271.6080.016-0.287-0.224-0.294
INP CBDclos　①-0.034-0.034-0.0321.026-0.318-0.235-0.270
INP CBDclos　②-0.0261.4150.012-0.297-0.047-0.272
BAN CTL 　①-0.039-0.039-0.0361.274-0.314-0.230-0.271
BAN CTL 　②-0.0561.530-0.060-0.284-0.084-3.11
INP CTL　①-0.042-0.043-0.0421.134-0.235-0.181-0.296
INP CTL　②-0.042-1.488-0.030-0.288-0.109-0.295
E1010　①-0.041-0.040-0.0361.171-0.322-0.252
E1010　②-0.0301.563-0.061-0.2670.286-0.267
blank ①/② /-0.307-0.306/-0.305-0.307/-0.2710.306/

9/22

Liquid Culture

 Sassa, Miyazaki
Namemedium
9/21 liquid culture BAN-CBDcexLB(AMP)
9/15 liquid culture INP-ctlLB(AMP)
9/15 liquid culture INP-CBDcexLB(AMP)
9/15 liquid culture INP-CBDclosLB(AMP)
9/15 liquid culture BAN-scFv 3C5LB(AMP)
9/15 liquid culture INP-scFv 3C5LB(AMP)
9/19 liquid culture BAN-CBDclosLB(AMP)
9/15 liquid culture BAN-ctlLB(AMP)
9/16 transformation J23105-RFP LB(AMP)

9/23

SEM

 Miyazaki
numbersample(&microl)VLP(&microl)PBScentrifugal force
1INP scFv /252.50×3000g
2INP CTL /252.50×3000g
3BclA scFv /252.50×3000g
4BclA CTL /252.50×3000g
5BclA scFv /2502.5×3000g
6INP scFv /252.50×200g
7INP CTL /252.50×200g
8BclA scFv /252.50×200g
9BclA CTL /252.50×200g
10BclA scFv /2502.5×200g
1102.50×200g

9/23

RNA extraction

 Isoda
numbersample name
1BclA-CBD cex
2BclA-CBDclos
3INP-CBDcex
4INP-CBDclos
5BclA-scFv
6INP-scFv

9/23

growth carve
sample0h0.5h1h1.5h2h3h4h5h6h7h8h9h10h
9/19 Liquid culture BAN scFv①0.0030.0010.0000.0080.0150.0090.0130.0160.0150.0220.0620.0200.027
9/19 Liquid culture BAN scFv②0.0130.0120.0000.0000.0000.0000.0010.0110.0110.0150.0220.0270.057
9/19 Liquid culture INP scFv①0.0120.0000.0000.0110.0060.0060.0070.0080.0130.0250.0360.0980.076
9/19 Liquid culture INP scFv②0.0070.0050.0000.0020.0030.0020.0120.0120.0190.0290.0340.0470.072
9/19 Liquid culture BAN cex①0.0140.0050.0100.0080.0110.0090.0140.0140.0160.0290.0380.0430.083
9/19 Liquid culture BAN cex②0.0110.0110.0100.0080.0040.0040.0200.0210.0360.0470.0650.0870.109
9/19 Liquid culture BAN clos①0.0000.0000.0000.0020.0090.0030.0090.0140.0190.0140.0160.0240.046
9/19 Liquid culture BAN clos②0.0050.0070.0000.0000.0000.0030.0120.0220.0200.0350.0550.0860.117
9/19 Liquid culture INP cex①0.1870.0000.0010.0070.0110.0060.0130.0160.0200.0230.0370.0480.068
9/19 Liquid culture INP cex②0.0020.0090.0000.0010.0010.0020.0150.0170.0230.0360.0530.0800.111
9/19 Liquid culture INP clos①0.0060.0000.0000.0230.0110.0030.0110.0130.0160.0220.0330.0580.068
9/19 Liquid culture INP clos②0.0290.0280.0000.0090.0000.0160.0010.0070.0080.0140.0200.0290.074
9/19 Liquid culture BAN CTL①0.0040.0000.0000.0000.0020.0000.0020.0120.0150.0120.0270.0550.081
9/19 Liquid culture BAN CTL②0.0020.0020.0010.0000.0010.0050.0170.0240.0310.0420.0590.0730.113
9/19 Liquid culture INP CTL①0.0110.0000.0320.0130.0170.0100.0150.0330.0310.0350.0370.0190.026
9/19 Liquid culture INP CTL②0.0130.0200.0000.0010.0000.0030.0090.0160.0210.0260.0440.0590.136
9/19 Liquid culture E1010①0.0150.0160.0140.0110.0160.0180.0320.0660.1080.1730.2580.3610.393
9/19 Liquid culture E1010②0.0640.0100.0070.0110.0120.0190.0290.0410.0610.0970.1500.180

9/23

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
11kb ladder/5
29/21 OEPCR/25

9/23

OEPCR

 Uchino
sample1 /&microlsample2 /&microlprimer Fw /&microlprimer Rv /&microlmilliQ /&microlKAPA HiFi /&microltotal
9/21 Gel Extract1 /19/21 Gel Extract2 /0.5VF/ 0.75VR /0.759.512.525

9/23

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
11kb ladder/3
29/23 OEPCR1 /25
39/23 OEPCR2 /25

9/23

PCR (Target)

 Yamamoto
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/8 miniprep INP-His-scFv9 /0.4m.scFv Fw/1.25VR/1.25KAPA2GFAST/12.59.6025
2/8 miniprep INP-His-scFv9 /0.4m.scFv Fw/1.25VR/1.25KAPA2GFAST/12.59.6025
2/8 miniprep INP-His-scFv9 /0.4VF/1.25m.scFv Rv/1.25KAPA2GFAST/12.59.6025
2/8 miniprep INP-His-scFv9 /0.4VF/1.25m.scFv Rv/1.25KAPA2GFAST/12.59.6025
9/23 Gel Extraction INP-scFv m.PCR/1.5VF/1.25VR/1.25KAPA2GFAST/12.58.5025

9/26

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
11kb ladder/4
29/23PCR1 /25
39/23PCR2 /25
49/23PCR3 /25
59/23PCR4 /25
69/23PCR5 /25

9/26

Gel Extraction

 Uchino
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
9/26 electrophoresis lane 4,50.357g
9/26 electrophoresis lane 2,30.354g

9/26

PCR (Target)

 Uchino
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
9/26 Gel Extract lane4,5 /2VF/0.75VR/0.752×KAPA HiFi Hotstart ReadyMix/12.5725
9/26 Gel Extract lane2,3 /2VF/0.75VR/0.752×KAPA HiFi Hotstart ReadyMix/12.5725

9/26

Liquid Culture

 Ohara
Namemedium
INP-scFv/30mlLB
INP-CTL/30mlLB
BAN-scFv/30mlLB
Ban CTL/30mlLB

9/26

Transformation

 Yamada
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
Bcla-cex 8/5 miniprepBL21LB(CP)
Bcla-CTL 8/4 miniprepBL21LB(CP)
Bcla-cex 8/5 miniprepBL21LB(CP)
Bcla-CTL 8/4 miniprepBL21LB(CP)

9/26

RNA extraction

 Isoda
numbersampleconc.&microg/&microl260/280260/230
4BclAcex74.731.941.61
2INPcex831.971.38
5INPclos120.211.991.94
3INPclos58.231.981.84
6BclAscFV74.761.961.84
7INPscFv80.61.971.97

9/27

Liquid Culture

 Yamada
Namemedium
BclA-cex /250mlLB(CP)
BclA-CTL /250mlLB(CP)

9/27

growth carve

 Sassa, Takimoto
sample0h0.5h1h1.5h2h3h4h5h6h7h8h
9/21Liquid culture BAN scFv0.1130.1700.2660.4080.5990.8930.164(1.64)0.169(1.69)0.232(2.32)0.234(2.34)0.221(2.21)
9/22Liquid culture BAN cex0.1090.1470.2190.3740.5150.8730.146(1.46)0.160(1.60)0.176(1.76)0.231(2.31)0.260(2.60)
9/19Liquid culture BAN clos0.0970.1090.1840.3380.4860.7440.130(1.30)0.173(1.73)0.199(1.99)0.203(2.03)0.274(2.74)
9/21Liquid culture BAN CTL0.1060.1700.2520.3040.5530.7970.177(1.77)0.155(1.55)0.216(2.16)0.236(2.36)0.257(2.57)
9/21Liquid culture INP scFv0.0880.0830.1010.1220.2990.4090.073(0.73)0.121(1.21)0.140(1.40)0.143(1.43)0.195(1.95)
9/15Liquid culture INP cex0.1020.1160.1450.2070.3200.6260.170(1.70)0.143(1.43)0.176(1.76)0.188(1.88)0.238(2.38)
9/22Liquid culture INP clos---0.1330.2090.3730.5720.9300.156(1.56)0.201(2.01)0.246(2.46)0.237(2.37)0.293(2.93)
9/21Liquid culture INP CTL0.1120.1550.2370.4090.5820.9550.153(1.53)0.209(2.09)0.268(2.68)0.288(2.88)0.274(2.74)
9/21Liquid culture E10100.0730.0910.1490.2650.3960.7060.157(1.57)0.182(1.82)0.222(2.22)0.222(2.22)

rpm 160
total10mL
CP　2mL/1L
4h~ 10倍希釈

9/27

Western Blotting

 Ohara
lanesample
MarkerM.Marker(Wako)
1BclA6His-scFv 300 &microl＋supernatant of NoVLP 3 &microl (washed by PBS)
2BclA6His ctl 300 &microl＋supernatant of NoVLP 3 &microl (NC) (washed by PBS)
3INPNC6His-scFv 300 &microl＋supernatant of NoVLP 3 &microl (washed by PBS)
4INPNC6His ctl 300 &microl＋supernatant of NoVLP 3 &microl (NC) (washed by PBS)
5BclA6His-scFv 300 &microl＋supernatant of NoVLP 3&microl (washed by PBST (0.1% tween20))
6BclA6His ctl 300 &microl＋supernatant of NoVLP 3&microl (NC) (washed by PBST (0.1% tween20))
7INPNC6His-scFv 300 &microl＋supernatant of NoVLP 3&microl (washed by PBST (0.1% tween20))
8INPNC6His ctl 300 &microl＋supernatant of NoVLP 3&microl (NC) (washed by PBST (0.1% tween20))
9PBS 300&microl＋supernatant of NoVLP 3 &microl (NC) (washed by PBS)

Transfer ;60 min 100 mA(CC)
Blocking ;1h
1’antibody ;anti-NoV pAb(mouse)
1:1000 with blocking buffer O/N
2’antibody ;anti-mouse HRP　
1:2500 with blocking buffer 1 h
Detection time　60 sec
(ECL Prime)

9/27

Western Blotting

 Ohara, Sassa
lanesample
MarkerM.Marker(Wako) 5&microl
1BBa_E1010(NC)
2BclA-6His-CBDclos (17 kDa)
3INPNC-6His-CBDclos(47 kDa)
4BclA-6His-CBDcex (17 kDa)
5INPNC-6His-CBDcex (47 kDa)
6BclA-6His-scFv (33 kDa)
7INPNC-6His-scFv (63 kDa)
8BBa_K875004 (PC) Lpp-OmpA-scFv-6His (43kDa)
Marker M.Marker(Wako) 3&microl
9BBa_K875004 (PC) Lpp-OmpA-scFv-6His(43 kDa)

Used membrane fraction from 100ml culture at logarithmic growth phase.

Transfer ;60min 100mA(CC)
Blocking ;1h
1’antibody ;anti6His(mouse)
1:1000 with blocking buffer O/N
2’antibody ;anti-mouse HRP　
1:2500 with TBST 1h
Detection time　380sec
(ECL Prime)

9/27

Reverse Transcription

 Yamada
numberTemplate/&microlgDNA wipeout buffer/&microlRNAase free water/&microltotal/&microl
A1BclAcex/122014
A2BclAclos/122014
A3INPcex/8.3223.6814
A4INPclos/122014
A5BclAscFv/122014
A6INPscFv/122014
B1BclAcex/122014
B2BclAclos/122014
B3INPcex/8.3223.6814
B4INPclos/122014
B5BclAscFv/122014
B6INPscFv/122014

9/27

Reverse Transcription

 Yamada
numbertemplate/&microlRTase/&microlRNAase free water/&microlRT buffer/&microlPrimer Mix/&microlTotal
A1BclAcex/14104120
A2BclAclos/14104120
A3INPcex/14104120
A4INPclos/14104120
A5BclAscFv/14104120
A6INPscFv/12104120
B1BclAcex/14014120
B2BclAclos/14014120
B3INPcex/14014120
B4INPclos/14014120
B5BclAscFv/14014120
B6INPscFv/12014120

9/27

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
11kb ladder/5
29/26 PCR INP- His scFv/25
39/26 PCR INP- His scFv/25

9/27

Gel Extraction

 Uchino
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
9/27 electrophoresis lane2,30.7660g

nanodrop
17.8ng/nl
260/280 1.79
260/230 0.31

9/27

CBD

 Yamada
15thnullnullnull
samplebeforeafterwash
BclAcex+ (CB buffer)0.181/0.212/0.2040.254/0.255/0.2540/0/0
BclAcex- (CB buffer)0.208/0.275/0.2280.160/0.171/0.162
BclActl+ (CB buffer)0.178/0.176/0.1710.082/0.085/0.0820.022/0.02/0.02
BclActl- (CB buffer)0.218/0.214/0.2160.161/0.160/0.159
BclAcex+ (PBS buffer)0.212/0.211/0.2100.333/0.334/0.3330.022/0.022/0.021
BclAcex- (PBS buffer)0.203/0.202/0.2010.178/0.178/0.180
BclActl+ (PBS buffer)0.182/0.181/0.1800.253/0.246/0.2460.021/0.021/0.021
BclActl- (PBS buffer)0.208/0.205/0.2080.192/0.194/0.194

9/29

PCR (Target)

 Uchino
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
9/27 Gel Extract/2VF/0.75VR/0.752×KAPA HiFi Hotstart ReadyMix/12.5925
VF/0.75VR/0.752×KAPA HiFi Hotstart ReadyMix/12.5925

9/29

Liquid Culture

 Ohara
Namemedium
BAN scFvLB
INP closLB
BAN closLB
INP cexLB
BAN cexLB
K875004LB

9/29

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
11kb ladder/2
29/29 PCR INP-His-scFv/2
3

9/29

PCR (Purification)

 Uchino
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
9/29 PCR INP-His-scFv31.92.021.38

9/30

Electrophoresis

 Uchino, Miyazaki
Lanesample/(&microl)Length(bp)
11kb ladder
2BclA clos/4.0
3INP clos/5.5
4BclA cex/4.5
5INP cex/2.5
6BclA scFv/4.5
7INP scFv/4.0

9/5

Restriction Enzyme Digestion

 Miyazaki
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
8/10 J23105 RFP miniprep 2010110×H 332

9/5

Transformation

 Yamamoto
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
8/29 Ligation Bclascfv-BclaCBDcex2CH5a/20LB(CP)
〃Bclascfv-BclaCBDclos2〃〃
〃Bclascfv-INPCBDcex2〃〃
〃Bclascfv-INPCBDclos2〃

9/5

Electrophoresis

 Yamamoto
Lanesample/(&microl)Length(bp)
18/20 SPcut Bcla-CBDcex/252520
2〃Bcla-CBDclos/25〃
3〃INP-CBDcex/25〃
4〃INP-CBDclos/25

9/5

Electrophoresis

 Miyazaki
Lanesample/(&microl)Length(bp)
1100bp ladder/56×loading Dye -
28/10 miniprep 9/5 xpcut J23105RFP/30

10/1

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
13RT- target /10
23RT+ target /10
33RT- 16s rRNA /10
43RT+ 16s rRNA /10
54RT- target /10
64RT+ target /10
74RT- 16s rRNA /10
84RT+ 16s rRNA /10
9100bp ladder /5
10A-6 target /10
11A-5 target /10
12A-6 16s rRNA /10
13A-5 16s rRNA /10
14B-6 target /10
15B-5 target /10
16B-6 16s rRNA /10
17B-5 16s rRNA /10

10/1

Transformation

 Ohara
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
K165002DH5α
BclA scFvDH5α
INP scFvDH5α
BclA cexDH5α
INP cexDH5α
BclA closDH5α
INP closDH5α
K165002BL21
BclA scFvBL21
INP scFvBL21
BclA cexBL21
INP cexBL21
BclA closBL21
INP closBL21

10/4

Transformation

 Yamada
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
INP CTL 8/4 miniprepDH5α/20LB(CP)

10/5

Liquid Culture

 Yamada
Namemedium
INP CTL 10/4 transformation

10/5

Western Blotting

 Ohara
lanesample
MarkerM.Marker(Wako) 5&microl
1BBa_E165002(NC)
2BclA-6His-scFv (33 kDa)
3INPNC-6His-scFv (63 kDa)
4BclA-6His-CBDcex (17 kDa)
5INPNC-6His-CBDcex(47kDa)
6BclA-6His-CBDclos (17 kDa)
7INPNC-6His-CBDclos (47 kDa)
8BBa_K875004(PC) Lpp-OmpA-scFv-6His (43 kDa)

Used pelet from 400 &microl overnight culture.

Transfar ;80 min 200 mA(CC)
Blocking ;O/N
1’antibody ;anti-6His(mouse)
1:1000 with blocking buffer 1h
2’antibody ;anti-mouse HRP　1:2500 with TBST 1h
Detection time　240 sec
(ECL Prime)

10/6

Liquid Culture

 Uchino
Namemedium
10/5 liquid culture INP CTL /2.5mlLB(CP) /47.5ml
10/5 liquid culture INP CTL /2.5mlLB(CP) /47.5ml
10/5 liquid culture INP CBD cex(5) /2.5mlLB(CP) /47.5ml
10/5 liquid culture INP CBD cex(5) /2.5mlLB(CP) /47.5ml

10/6

Liquid Culture

 Yamada
Namemedium
10/5 liquid culture INP CTL /100mlLB(CP) /100ml
10/3 liquid culture INP CBD cex /100mlLB(CP) /200ml

10/7

CBD

 Yamada
16thnullnullnull
samplebeforeafterwash
INPcex+0.208/0.208/0.2110.224/0.221/0.2260.013/0.012/0.010
INPcex-0.210/0.216/0.2040.232/0.231/0.232
INPctl+0.196/0.196/0.1910.220/0.217/0.2120.023/0.024/0.025
INPctl-0.194/0.195/0.1960.207/0.199/0.208
cellulose only0.015/0.013/0.0150.031/0.031/0.0320.018/0.017/0.019

10/11

Transformation

 Yamada
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
2/25 miniprep INPcex3DH5α /20LB(CP)×3
2/8 miniprep INP His 12 (INPscFv)3DH5α /20LB(CP)×3
2/25 miniprep INPcex3BL21 /20LB(CP)×3
2/8 miniprep INP His 12 (INPscFv)3BL21 /20LB(CP)×3

10/11

SEM(CBD)

 Miyazaki
sample/mlbeforeafterwash1wash2
INP CBDcex 10/1 liquid culture /900.5500.3920.074/0.084-0.004/-0.018
INP CBDcex 10/1 liquid culture /450.5500.4320.002/0.000-0.001/0.000

10/12

Colony PCR (typeA)

 Yamada
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
INP-His-scFv-INP-His-CBDcex DH5α colony1VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony2VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony3VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony4VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony5VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony6VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony7VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony8VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony9VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony10VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony11VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony12VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony13VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony14VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony15VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony16VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony17VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony18VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony19VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony20VF/0.5 VR/0.511.5Quick Taq/12.525

10/12

Colony PCR (typeA)

 Yamada
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
INP-His-scFv-INP-His-CBDcex DH5α colony21VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony21VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony21VF/0.5 VR/0.511.5Quick Taq/12.525
INP-His-scFv-INP-His-CBDcex DH5α colony21VF/0.5 VR/0.511.5Quick Taq/12.525
2/8 miniprep INP-His-scFv /0.5VF/0.5 VR/0.511.5Quick Taq/12.525
2/25 miniprep INP-His-CBDcex /0.5VF/0.5 VR/0.511.5Quick Taq/12.525

10/12

Electrophoresis

 Yamada
Lanesample/(&microl)Length(bp)
1INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony1 /5
2INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony2 /5
3INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony3 /5
4INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony4 /5
5INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony5 /5
6INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony6 /5
7INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony7 /5
8INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony8 /5
9INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony17 /5
10INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony18 /5
11INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony19 /5
12INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony20 /5
13INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony21 /5
14INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony22 /5
15INPNC-His-scFv-INPNC-His-CBDcex 10/12 colony PCR colony23 /5
16INPNC-His-CBDcex 10/12 colony PCR /5
171kbp ladder /10

10/12

PCR (Target)

 Uchino
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/8 INP-His 12 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025

10/13

PCR (Target)

 Uchino
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/8 INP-His 12 /0.1VF /0.75VR /0.752×KAPA /12.510.9025

10/13

Colony PCR

 Yamada
sample /&microlprimer /&microlKAPA Taq Extra Hotstart /&microlDMSO /&microlMilliQ /&microltotal /&microl
INP-cex + INP-scFv colony2VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony7VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony17VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony19VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony9VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony10VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony11VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony12VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony13VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony16VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony23VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony24VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony25VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony26VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony27VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony28VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony29VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony30VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony31VF /1.25 VR /1.2512.50.759.2525

10/13

Colony PCR

 Yamada
sample /&microlprimer /&microlKAPA Taq Extra Hotstart /&microlDMSO /&microlMilliQ /&microltotal /&microl
INP-cex + INP-scFv colony31VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony32VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony33VF /1.25 VR /1.2512.50.759.2525
INP-cex + INP-scFv colony34VF /1.25 VR /1.2512.50.759.2525
INP-scFv /0.25VF /1.25 VR /1.2512.50.759.2525
INP-cex /0.25VF /1.25 VR /1.2512.50.759.2525

10/13

Miniprep

 Uchino
DNAConcentration/(&microg/ml)260/280260/230
10/6 liquid culture INP-His-CBDcex27.81.961.86

10/13

PCR (Purification)

 Uchino
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
10/12 PCR INP-His 12 (INP-His coding)727.31.881.86
10/13 PCR INP-His 121132.11.892.04

10/13

Restriction Enzyme Digestion

 Uchino
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
10/13 PCR purification INP-His coding7200210×H /604460
10/13 PCR purification INP-His 126020210×M /604460
10/13 midiprep INP-His-CBDcex50200210×H /60060
10/13 midiprep INP-His-CBDcex50002210×H /60060

10/13

Electrophoresis

 Yamada
Lanesample/(&microl)Length(bp)
1INPcex+INPscFv colony2
2INPcex+INPscFv colony7
3INPcex+INPscFv colony19
4INPcex+INPscFv colony9
5INPcex+INPscFv colony17
6INPcex+INPscFv colony10
7INPcex+INPscFv colony11
8INPcex+INPscFv colony12
9INPcex+INPscFv colony15
10INPcex+INPscFv colony16
11INPcex+INPscFv colony23
12INPcex+INPscFv colony24
13INPcex+INPscFv colony25
14INPcex+INPscFv colony26
15INPscFv /5
16INPcex /5
171kbp ladder

10/13

Electrophoresis

 Yamada
Lanesample/(&microl)Length(bp)
A1INPcex+INPscFv colony2
A2INPcex+INPscFv colony7
A3INPcex+INPscFv colony19
A4INPcex+INPscFv colony9
A5INPcex+INPscFv colony17
A6INPscFv /5
A7INPcex /5
A81kbp ladder /5
B1INPcex+INPscFv colony10
B2INPcex+INPscFv colony11
B3INPcex+INPscFv colony12
B4INPcex+INPscFv colony13
B5INPcex+INPscFv colony14
B61kbp ladder /5

10/13

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
11kbp ladder
210/13 1A
310/13 2A
410/13 3A
510/13 4A
610/13 5A
710/13 1B
810/13 2B
910/13 3B
1010/13 4B
1110/15 5B

10/13

Gel Extraction

 Uchino
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
1A+2A10/13 PCR purification INP His coding
3A~5A10/13 midiprep INP His CBDcex
1B+2B10/13 PCR purification INP His12
3B~5B10/13 midiprep INP His CBDcex

10/13

Fluorescence Microscopy

 Uchino
samplenumbermagnification
INPclos13318×200
INPclos13319~13328×600
INP CTL13329×200
INP CTL13330~13338×600
cellulose only13339×200
cellulose only13340,13343~13351×600
cellulose only13341,13342×1000

without filter

10/13

Fluorescence Microscopy

 Uchino
E.coliOD600celluloseweightfilter
--powder10non-filtered
INPNC-His-ctl0.2powder10non-filtered
INPNC-His-CBDcex0.2powder10non-filtered
--powder10filtered
INPNC-His-ctl0.2powder10filtered
INPNC-His-CBDcex0.2powder10filtered

10/13

Fluorescence Microscopy

 Uchino
samplenumbermagnification
INPcex13353×200
INPcex13354~13356,13364,13366×600
INPcex13357,13365×1000
INP CTL13367×200
INP CTL13368~13378×600
INP CTL13379×1000
cellulose only13380,13381×200
cellulose only13382~13391×600
cellulose only

with filter

10/13

Fluorescence Microscopy

 Uchino
samplenumbermagnification
INPcex13392~98×200
INPcex13399~13408×600
INP CTL13309~13419×200
bemliese13420~13430×200

with bemliese

10/14

Electrophoresis

 Yamamoto
Lanesample/(&microl)Length(bp)
11kbp ladder /3
210/13 1A /25
410/13 2A /25
510/13 3A /25
610/13 4A /25
710/13 5A /25
810/13 1B /25
910/13 2B /25
1010/13 3B /25
1110/13 4B /25
1210/13 5B /25

with bemliese

10/14

SEM(CBD)

 Miyazaki
samplemembrane shape
INPcex + cellulosesquare
INP CTL + cellulosetriangle
cellulose onlypentagon

10/14

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
110/14 gel extract e2 /5

10/14

Gel Extraction

 Uchino
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
10/14 electrophoresis 1B longer + 2B longer e110/13 restriction enzyme design INP His 12
10/14 electrophoresis 1B longer + 2B longer e210/13 restriction enzyme design INP His12

10/14

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
11kbp ladder /3
21A + 2A /3
33A + 4A + 5A /3

10/14

Gel Extraction

 Uchino
LaneRestriction Enzyme Digestion ProductVolume/(&microl)
10/14 electrophoresis 1B + 2B10/13 restriction enzyme design INP His12
10/14 electrophoresis 1A + 2A10/13 restriction enzyme design INP His coding
10/14 electrophoresis 3A + 4A + 5AINP His CBDcex

10/14

Ligation

 Uchino
sample1 1A + 2A /&microlsample2 3A~5A /&microlLigation high ver.2total
52.57.515
3.753.757.515

10/14

Transformation

 Uchino
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
INP His coding ligationA2DH5α /20LB(CP)
INP His coding ligationA4DH5α /20LB(CP)
INP His coding ligationB2DH5α /20LB(CP)
INP His coding ligationB4DH5α /20LB(CP)

10/15

PCR (Purification)
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
10/15 INP-His coding i207.41.872.11
10/15 INP-His coding G174.71.881.09
10/15 INP-His coding E175.51.082.10
10/15 INP-His coding M212.91.88

10/15

CBD　Binding Assay

 Ohara
sampleBufferOD
INP cexCBC buffer1.002
simultaneous expressionCBC buffer0.994
INP CTLCBC buffer1.004
INP CTLPBS0.964
simultaneous expressionPBS

10/15

Restriction Enzyme Digestion
SampleDNA/(&microl)EcoR1/(&microl)Xba1/(&microl)Spe1/(&microl)Pst1/(&microl)Buffer/(&microl)BSA/(&microl)MilliQ/(&microl)Total/(&microl)
10/15 PCR purification INP His coding i29.0200262160
10/15 PCR purification INP His coding G34.32002615.760
10/15 PCR purification INP His coding E34.22002615.860
10/15 PCR purification INP His coding M28.2(6000ng)2002621.860
10/13 midiprep INP-His-CBDcex5020026060
10/13 midiprep INP-His-CBDcex5020026060
10/13 midiprep INP-His-CBDcex5020026060
10/13 midiprep INP-His-CBDcex502002060

1.5　Dr.Kitabatake
2.6 Agarose S ＋　Promega Gel Extraction
3.7 Agarose S ＋　Dr.Nasuda
4.8　stock

10/15

Gel strecking

 Yamada
samplenull
INPNc-His-scFv (CP)(LB)plate×3
INP-His-CBDcex

10/15

PCR (Target)

 Uchino
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025
2/22 INP-His scFv colony3 /0.1INP His coding Fw /0.75INP His coding Rv /0.752×KAPA /12.510.9025

10/15

Colony PCR (typeA)

 Yamamoto
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
A1 10/14 INPcex + INPscFv colony1VF/1.25 VR/1.258.75KAPA2G /12.5 DMSO/1.2525
A2 10/14 INPcex + INPscFv colony2VF/1.25 VR/1.258.75KAPA2G /12.5 DMSO/1.2525
A3 10/14 INPcex + INPscFv colony3VF/1.25 VR/1.258.75KAPA2G /12.5 DMSO/1.2525
A4 10/14 INPcex + INPscFv colony4VF/1.25 VR/1.258.75KAPA2G /12.5 DMSO/1.2525

10/15

Western Blotting

 Ohara
lanesample
MarkerM.Marker
1BBa_K875004(PC) Lpp-OmpA-scFv-6His 43kDa
2INPNC-6His-CBDcex 47kDa
3INPNC-6His-scFv 63kDa
4INPNC-6His-CBDcex/INPNC-6His-scFv
5BBa_K165002(NC)

Used pelet from 500 &microl overnight culture, adjested OD600 to 3.1 .

Transfar ;80 min 200 mA(CC)
Blocking ;O/N
1’antibody ;anti-6His(mouse)
1:1000 with blocking buffer 1h
2’antibody ;anti-mouse HRP　1:2500 with TBST 1h
Detection time　900 sec
(ECL Prime)

10/16

Electrophoresis

 Yamamoto
Lanesample/(&microl)Length(bp)
1kb ladder
10/15 colony PCR colony 13
10/15 colony PCR colony 225
10/15 colony PCR colony 325
10/15 colony PCR colony 4

10/16

Cellulose binding Assay

 Uchino
Cellulose/(mg)E.coli/(mg)buffer
1cex/1carbonate-bicarbonate buffer
1ctl/1carbonate-bicarbonate buffer
0.5cex/1carbonate-bicarbonate buffer
0.5ctl/1carbonate-bicarbonate buffer
0.5CBC/1carbonate-bicarbonate buffer
0.1cex/1carbonate-bicarbonate buffer
0.1ctl/1

RT,-3h 500rpm 15:45~21:15 binding
fix with 4×1/100 PFA 10min→centrifuge→PBS→centrifuge
→PBS→centrifuge→PBS→centrifuge→DAP1→centrifuge
→PBS→centrifuge→PBS→through filter→PBS into filter
→centrifuge→PBSinto filter→centrifuge
→suspend with PBS→centrifuge
→suspend with?discoloration inhibitor

10/16

Dual Display Assay for Electron Microscope
E.coliCelluloseVLPbuffer
A.Dual 1mL1mg10&microLPBS
B.Dual 1mL1mgPBS
C.Dual 1mL10&microLPBS
D.Dual 1mLPBS
E.INPctl1mg10&microLPBS
F.INPctl1mgPBS
G.INPctl10&microLPBS
H.INPctlPBS
I1mgPBS
J10&microL

RT,ih incubate,850rpm
Dul=1.020
ctl=1.056

10/16

Electrophoresis
Lanesample/(&microl)Length(bp)
11 kb ladder5
210/15 Digestion INP His Coding 230 /0.94 上0.04下0.10
310/15 Digestion INP His Coding 230 /0.94 上0.04下0.19
410/13 miniprep INP-His-CBDcex 630 /0.94 0.11
510/13 miniprep INP-His-CBDcex 630 /0.94

Uchino 16:08~
ABEF→centrifuge→resupend with PBS →centrifuge→through filter→resupend with PBS→through filter→centrifuge→respend with PBS→centrifuge→resupend with PBS→centrifuge→resupend with PBS
IJ　Michimori

10/16

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
11kb ladder
210/15 colony PCR colony1
310/15 colony PCR colony2
410/15 colony PCR colony3
510/15 colony PCR colony

10/16

PCR (Target)

 Sassa
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
2/22 scFv col.3 0.1VF 0.75VR 0.75KAPA HoFi 12.510.925
2/22 scFv col.3 0.1VF 0.75VR 0.75KAPA HoFi 12.510.9

95℃ 5min
98℃ 20s×35
55℃ 15s ×35
72℃ 30s×35
4℃ infinity

10/16

Miniprep

 Uchino, Yamamoto
DNAConcentration/(&microg/ml)260/280260/230
10/15 Gel Extraction INP His coding 長9.61.880.27
10/15 Gel Extraction INP His coding 短74.41.870.84
10/15 Gel Extraction INP His CBDcex22.01.970.33
Insert MK64.51.921.17
Vector MK56.62.37

10/16

Ligation

 Yamamoto
InsertVectorMilliQLigation High
10/15 Insert MK/2&microL10/15 Gel Extract INP His CBDcex 5.5&microl07.5
10/15 Insert MK/1&microL10/15 Gel Extract INP His CBDcex 6&microL07.5
10/15 Insert MK/1&microL10/15 Gel Extract INP His CBDcex 0.5&microL67.5
10/15 Digestion INP His coding/5&microL10/15 Digestion INP His CBDcex 2.5&microL07.5
10/15 Digestion INP His coding/6&microL10/15 Digestion INP His CBDcex 1.5&microL07.5
10/15 Digestion INP His coding/4.5&microL10/15 Digestion INP His CBDcex 3&microL07.5
10/14Gel Extract INP His coding/5.510/15 Digestion INP His CBDcex 2&microL07.5
10/14Gel Extract INP His coding/610/15 Digestion INP His CBDcex 1.5&microL07.5
10/15 Digestion INP His coding 3.5&microL10/15 Digestion INP His CBDcex 4&microL07.5
10/15 Digestion INP His coding 2.5&microL10/15 Digestion INP His CBDcex 5&microL07.5
10/15 Digestion INP His coding 3.7510/15 Digestion INP His CBDcex 3.750

10/16

Transformation

 Yamamoto
Sample NameSample Volume (&microl)Competent Cells/(&microl)Medium
10/15Ligation Insert MK＋Gel Extract1 2&microL2DH5α/2
10/15Ligation Insert MK＋Gel Extract2 2&microL2DH5α/2
10/15Ligation Digestion＋Digestion3 2&microL2DH5α/2
10/15Ligation Digestion＋Digestion4 2&microL2DH5α/2
10/15Ligation Digestion＋Digestion5 2&microL2DH5α/2
10/15Ligation Digestion＋Digestion6 2&microL2DH5α/2
10/15Ligation Gel Extract＋Digestion7 2&microL2DH5α/2
10/15Ligation Gel Extract＋Digestion8 2&microL2DH5α/2
10/15Ligation Digestion＋Digestion9 2&microL2DH5α/2
10/15Ligation Digestion＋Digestion10 2&microL2DH5α/2
10/15Ligation Digestion＋Digestion11 2&microL2DH5α/2
10/14 Ligation PuroductA 2&microL2DH5α/2
10/14 Ligation PuroductB 2&microL2

10/16

PCR (Purification)

 Yamamoto
Sample Name/(&microl)Concentration(ng/&microl)260/280260/230
10/15 PCR scFv INP His codingFw＋INP His RV284.41.88

10/16

Liquid Culture

 Yamamoto
Namemedium
10/14 Tranceformation INPHis coding2 colony1LB(CP)
10/14 Tranceformation INPHis coding2 colony2

10/16

non-woven felt

Ohara
samplenull
cex13431×200
32~41×600
ctl13431×200

10/16

Liquid Culture

 Yamamoto
Namemedium
10/14 Trance formation INP His coding 2 colony1LB(CP)
10/14 Trance formation INP His coding 2 colony2

10/16

fluorescence microscope assay

 Uchino
samplenumbermagnification
cex bemliese13431×200
cex bemliese13432~13441×600
CTL bemliese13442×200
CTL bemliese13443~13355×600

10/16

Colony PCR

 Matsumoto, Uchino
numbersampleAny othersAny others(&microL)
1~510/15Ligation Insert MK＋Gel Extract12×Ready Mix12.5
6~1010/15Ligation Insert MK＋Gel Extract2Fwd primer1.25
11~1510/15Ligation Insert MK＋Gel Extract3Rev primer1.25
16~2010/15Ligation Digestion＋Digestion4MilliQ10
21~2410/15Ligation Digestion＋Digestion552×mix
25~2810/15Ligation Digestion＋Digestion62×Ready650
29~3210/15Ligation Gel Extract＋Digestion7Fwd 65
33~3610/15Ligation Gel Extract＋Digestion8Rev65
37~4010/15Ligation Digestion＋Digestion9MilliQ
41~4410/15Ligation Digestion＋Digestion10
45~4810/15Ligation Digestion＋Digestion11

×16
16Colony Picking　DMSO 0.75 12.75 12
Mix 12.5 212.5 200
Fwd 1.25 21.25 18.75 20
Rev 1.25 21.25 18.75 20
MilliQ 9.25 157.25 138.75 148
16sample colony1~14・・・P15のsample colony
Blue　　　　　　　15・・・2/8 INP-His9
16・・・8/1 miniprep INPcex colony

10/16

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
11kb ladder
21
32
43
54
65
76
87
98
109
11
12

10/16

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
1
21kb ladder
311
412
513
614
715
816
917
1018
1119
1220

10/16

Fluorescence Microscopy

 Uchino
E.coliOD600cellulosesize / weightfilter
--bemliese5mm×5mm-
INPNC-His-ctl1bemliese5mm×5mm-
INPNC-His-CBDcex1bemliese5mm×5mm-
--powder1filtered
INPNC-His-ctl1powder1filtered
INPNC-His-CBDcex1powder1filtered
--powder0.5filtered
INPNC-His-ctl1powder0.5filtered
INPNC-His-CBDcex1powder0.5filtered
--powder0.1filtered
INPNC-His-ctl1powder0.1filtered
INPNC-His-CBDcex1powder0.1filtered

10/16

fluorescence microscope assay

 Uchino
samplenumbermagnification
cex 1mg13456×200
cex 1mg13457~13466×600
cex 0.5mg13467×200
cex 0.5mg13468~13378×600
cex 0.5mg13379×1000
CTL 1mg13380×200
CTL 1mg13381~13390×600
CTL 0.5mg13391×200
CTL 0.5mg13392~13401×600
CTL 0.5mg13402×1000
cex 0.1mg13403×200
cex 0.1mg13404,13407~13416×600
cex 0.1mg13405,13406×1000
CTL 0.1mg13417×200
CTL 0.1mg13418~13430×600
CTL 0.1mg13431×200
cellulose 0.5mg13432~13441×600

10/17

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
121
222
31kb ladder
423
524
625
726
827
928
1029
11
12

10/17

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
11kb ladder
231
332
433
534
6

10/17

Electrophoresis
Lanesample/(&microl)Length(bp)
136
237
338
439
5
6

10/17

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
11kb ladder
241
342
443
5
6

10/17

Electrophoresis

 Uchino
Lanesample/(&microl)Length(bp)
1
21kb ladder
345
446
547
6

10/17

Liquid Culture

 Yamamoto
Namemedium
colony 1,2,3,4,5,11,12,13,14,15,18,19,22,24,26,30

total 16mL

10/17

Miniprep

 Uchino
DNAConcentration/(&microg/ml)260/280260/230
165.81.91
263.21.901.84
384.61.871.33
4100.81.921.67
584.61.891.59
1189.31.841.43
1287.31.921.88
1388.01.891.59
1479.81.911.72
1583.31.901.82
18102.51.881.50
19114.91.911.59
22109.91.781.14
2480.41.932.00
2664.61.931.84
3088.81.801.07

total 16mL

10/17

Colony PCR (typeA)

 Yamada
Templates/(&microl)Primers/(&microl)MilliQ/(&microl)PreMix/(&microl)Total/(&microl)
10/15gel plated colony 1VF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
10/15gel plated colonyVF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
10/15gel plated colonyVF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
10/15gel plated colonyVF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
10/15gel plated colonyVF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
10/15gel plated colonyVF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
10/15gel plated colonyVF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
10/15gel plated colonyVF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
10/15gel plated colonyVF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
10/15gel plated colonyVF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
10/15gel plated colonyVF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
10/15gel plated colonyVF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
10/15gel plated colonyVF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
10/15gel plated colonyVF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
INPNC-His-scFvVF1.25 VR1.259.25KapaTag/12.5 DMSO 0.7525
INP-His-CBDcexVF1.25 VR1.259.2525

VF 1.25×8=10&microL
VR 1.25×8=10&microL
Kapa 12.5×8=100&microL
DM80 0.75×8=6&microL
MilliQ 9.25×8=74&microL
Total 200&microL

10/17

CBD Binding Assay

 Uchino
Dual 1mL OD=1.0bemlise 5mm×5mmVLP Narita104 10&microL
Dual 1mL OD=1.0bemlise 5mm×5mm
ctll 1mL OD=1.0bemlise 5mm×5mm
ctl 1mL OD=1.0bemlise 5mm×5mm

10/1

RNA purification

 Ohara
BclAcex0.504
BclAscFv0.788
INPscFv0.837
INPclos1.564
BclAclos0.817
INPcex0.839

10/1

RNA extraction

 Uchino
numbersampleconc(ng/&microl)260/280260/230diluted solution volume(200ng/&microl)
1INPclos359.22.071.2111.94
2BclAcex230.92.030.852.31
3INPcex1069.02.111.4365.17
4INPscFv320.02.100.939
5BclAclos386.22.040.9713.97
6BclAcex441.02.040.8318

10/1

Reverse Transcription

 Uchino
namecopy(high or low)RT
3 negahigh copy-
3 posihigh copy+
4 negalow copy-
4 posilow copy+

10/1

PCR (Target)

 Uchino
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
10/1 RT×20 3RT- /1.0&microlrRNA16s forward primer1 /0.4&microlrRNA16s reverse primer1 /0.4&microl2×KAPA HiFi /5&microl3.2&microl10&microl
10/1 RT×20 3RT+ /1.0&microlrRNA16s forward primer1 /0.4&microlrRNA16s reverse primer1 /0.4&microl2×KAPA HiFi /5&microl3.2&microl10&microl
10/1 RT×20 4RT- /1.0&microlrRNA16s forward primer1 /0.4&microlrRNA16s reverse primer1 /0.4&microl2×KAPA HiFi /5&microl3.2&microl10&microl
10/1 RT×20 4RT+ /1.0&microlrRNA16s forward primer1 /0.4&microlrRNA16s reverse primer1 /0.4&microl2×KAPA HiFi /5&microl3.2&microl10&microl
9/27 RT×1 A-6 /1.0&microlrRNA16s forward primer1 /0.4&microlrRNA16s reverse primer1 /0.4&microl2×KAPA HiFi /5&microl3.2&microl10&microl
9/27 RT×1 A-5 /1.0&microlrRNA16s forward primer1 /0.4&microlrRNA16s reverse primer1 /0.4&microl2×KAPA HiFi /5&microl3.2&microl10&microl
9/27 RT×1 B-6 /1.0&microlrRNA16s forward primer1 /0.4&microlrRNA16s reverse primer1 /0.4&microl2×KAPA HiFi /5&microl3.2&microl10&microl
9/27 RT×1 B-5 /1.0&microlrRNA16s forward primer1 /0.4&microlrRNA16s reverse primer1 /0.4&microl2×KAPA HiFi /5&microl3.2&microl10&microl

10/1

PCR (Target)

 Uchino
Templates/(&microl)Primer1/(&microl)Primer2/(&microl)Buffer/(&microl)MilliQ/(&microl)Any Other/(&microl)Total/(&microl)
10/1 RT×20 3RT- /1.0INP primer for CBD /0.4CBD cex primer Rv /0.42×KAPA HiFi /53.210
10/1 RT×20 3RT+ /1.0INP primer for CBD /0.4CBD cex primer Rv /0.42×KAPA HiFi /53.210
10/1 RT×20 4RT- /1.0scFv primer1 /0.4scFv primer2 /0.42×KAPA HiFi /53.210
10/1 RT×20 4RT+ /1.0scFv primer1 /0.4scFv primer2 /0.42×KAPA HiFi /53.210
9/27 RT×1 A-6 /1.0INP primer for CBD /0.4CBD cex primer Rv /0.42×KAPA HiFi /53.210
9/27 RT×1 A-5 /1.0INP primer for CBD /0.4CBD cex primer Rv /0.42×KAPA HiFi /53.210
9/27 RT×1 B-6 /1.0scFv primer1 /0.4scFv primer2 /0.42×KAPA HiFi /53.210
9/27 RT×1 B-5 /1.0scFv primer1 /0.4scFv primer2 /0.42×KAPA HiFi /53.210