Difference between revisions of "Team:Kyoto/Notebook"

(Undo revision 329740 by Hkaneko (talk))
(Undo revision 329685 by Hkaneko (talk))
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<a name="0821" class = "kyoto-jump"></a>
 
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<span class="keln_date"><h3>8/21</h3></span>
 
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<span class="keln_exp"><h4>Liquid Culture</h4></span>
 
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<span class="keln_researcher">Li</span>
 
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<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>
 
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<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>
 
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<tr><td>8/20 transformation p+ [1]</td><td>PG(AMP) /6</td></tr>
 
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<tr><td>8/20 transformation p- [1]</td><td>PG(AMP) /6</td></tr>
 
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</table>
 
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</div>
 
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<a name="0821" class = "kyoto-jump"></a>
 
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<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>
 
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<img src="https://static.igem.org/mediawiki/2016/9/9d/T--Kyoto--8_21_No34_Electrophoresis_Kaneko_05_20_05_13_01.jpg">
 
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</div>
 
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<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>
 
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<a name="0824" class = "kyoto-jump"></a>
 
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<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>
 
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</table>
 
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</div>
 
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<div class="keln_container">
 
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<a name="0824" class = "kyoto-jump"></a>
 
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<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>
 
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</table>
 
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</div>
 
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<a name="0824" class = "kyoto-jump"></a>
 
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<span class="keln_date"><h3>8/24</h3></span>
 
−
<span class="keln_exp"><h4>Miniprep</h4></span>
 
−
<span class="keln_researcher">Li</span>
 
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<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>
 
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</table>
 
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</div>
 
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<!------------ Table END ------------>
 
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<!-- Table Generated by KELN Author: Iguchi -->
 
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<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>
 
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</table>
 
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</div>
 
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<!-- Table Generated by KELN Author: Kaneko -->
 
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<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>
 
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</table>
 
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</div>
 
−
<!------------ Table END ------------>
 
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<!-- 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 ------------>
 
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<!-- 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>
 
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</div>
 
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<!------------ Table END ------------>
 
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<!-- Table Generated by KELN Author: Kaneko -->
 
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<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">
 
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</div>
 
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<p class="keln_note">[img42]</p>
 
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<!------------ Table END ------------>
 
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<!-- Table Generated by KELN Author: Kaneko -->
 
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<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>
 
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</table>
 
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</div>
 
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<!------------ 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>
 
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<!------------ 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">
 
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</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 1,491: Line 189:
 
</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>
+
 
−
<span class="keln_date"><h3>2/6</h3></span>
+
<a name="0101" class = "kyoto-jump"></a>
−
<span class="keln_exp"><h4>Miniprep</h4></span>
+
 
−
<span class="keln_researcher">Notsu, Kaneko, Egashira, Wan, Yamamoto</span>
+
<span class="keln_date"><h3>1/1</h3></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><td>2/5 liquid culture E1010(RFP)  colony1</td><td>48.0</td><td>1.93</td><td></td></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>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>
+
 
−
<span class="keln_date"><h3>2/6</h3></span>
+
<a name="0101" class = "kyoto-jump"></a>
−
<span class="keln_exp"><h4>Restriction Enzyme Digestion</h4></span>
+
 
−
<span class="keln_researcher">Kaneko, Egashira, Wan, Yamamoto</span>
+
<span class="keln_date"><h3>1/1</h3></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><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><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>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>
+
 
−
</table>
+
<tr><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
−
</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>
+
 
−
<span class="keln_date"><h3>2/15</h3></span>
+
<a name="0101" class = "kyoto-jump"></a>
−
<span class="keln_exp"><h4>Liquid Culture</h4></span>
+
 
−
<span class="keln_researcher">Wan, Yamamoto</span>
+
<span class="keln_date"><h3>1/1</h3></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><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><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>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>
+
 
−
<span class="keln_date"><h3>2/15</h3></span>
+
<a name="0101" class = "kyoto-jump"></a>
−
<span class="keln_exp"><h4>Electrophoresis</h4></span>
+
 
−
<span class="keln_researcher">Kaneko</span>
+
<span class="keln_date"><h3>1/1</h3></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><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><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-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>
+
 
−
<span class="keln_date"><h3>2/16</h3></span>
+
<a name="0101" class = "kyoto-jump"></a>
−
<span class="keln_exp"><h4>Gel Extraction(Measurement)</h4></span>
+
 
−
<span class="keln_researcher">Yamamoto</span>
+
<span class="keln_date"><h3>1/1</h3></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><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><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>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>
+
 
−
<span class="keln_date"><h3>2/16</h3></span>
+
<a name="0101" class = "kyoto-jump"></a>
−
<span class="keln_exp"><h4>Miniprep</h4></span>
+
 
−
<span class="keln_researcher">Uchino, Yamada</span>
+
<span class="keln_date"><h3>1/1</h3></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><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><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-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>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>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-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>
+
 
−
<span class="keln_date"><h3>2/17</h3></span>
+
<a name="0101" class = "kyoto-jump"></a>
−
<span class="keln_exp"><h4>Gel Extraction</h4></span>
+
 
−
<span class="keln_researcher">Michimori, Wan, Yamamoto, Kim</span>
+
<span class="keln_date"><h3>1/1</h3></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><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><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-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>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>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-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>
+
 
−
<span class="keln_date"><h3>2/17</h3></span>
+
<a name="0101" class = "kyoto-jump"></a>
−
<span class="keln_exp"><h4>Miniprep</h4></span>
+
 
−
<span class="keln_researcher">Matsumoto, Kim</span>
+
<span class="keln_date"><h3>1/1</h3></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><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><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-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>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>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</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>
+
 
−
<span class="keln_date"><h3>2/18</h3></span>
+
<a name="0101" class = "kyoto-jump"></a>
−
<span class="keln_exp"><h4>Colony PCR (typeA)</h4></span>
+
 
−
<span class="keln_researcher">Uchino</span>
+
<span class="keln_date"><h3>1/1</h3></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><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><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>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>J23114</td><td>VF/ 0.75</td><td>J23114+RBSRv</td><td></td><td>11</td><td></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="0226" class = "kyoto-jump"></a>
+
 
−
<span class="keln_date"><h3>2/26</h3></span>
+
<a name="0101" class = "kyoto-jump"></a>
 +
 
 +
<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><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><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>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>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>
+
<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="0226" class = "kyoto-jump"></a>
+
 
−
<span class="keln_date"><h3>2/26</h3></span>
+
<a name="0101" class = "kyoto-jump"></a>
−
<span class="keln_exp"><h4>Electrophoresis</h4></span>
+
 
−
<span class="keln_researcher">Egashira</span>
+
<span class="keln_date"><h3>1/1</h3></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><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><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>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>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>
+
<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>
+
 
−
<!------------ 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 END ------------>
+
<!-- Table Generated by KELN Author: Sukegawa -->
−
<!-- 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>
 
  
−
<!------------ Table END ------------>
+
<a name="0101" class = "kyoto-jump"></a>
−
<!-- Table Generated by KELN Author: Miyazaki -->
+
 
−
<div class="keln_container">
+
<span class="keln_date"><h3>1/1</h3></span>
−
<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 13:38, 18 October 2016

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

1/1

PCR (Target)

 Sukegawa
Templates/(μl)Primer1/(μl)Primer2/(μl)Buffer/(μl)MilliQ/(μl)Any Other/(μl)Total/(μl)
1111111

1/1

PCR (Target)

 Sukegawa
Templates/(μl)Primer1/(μl)Primer2/(μl)Buffer/(μl)MilliQ/(μl)Any Other/(μl)Total/(μl)
1111111

1/1

PCR (Target)

 Sukegawa
Templates/(μl)Primer1/(μl)Primer2/(μl)Buffer/(μl)MilliQ/(μl)Any Other/(μl)Total/(μl)
1111111

1/1

PCR (Target)

 Sukegawa
Templates/(μl)Primer1/(μl)Primer2/(μl)Buffer/(μl)MilliQ/(μl)Any Other/(μl)Total/(μl)
1111111

1/1

PCR (Target)

 Sukegawa
Templates/(μl)Primer1/(μl)Primer2/(μl)Buffer/(μl)MilliQ/(μl)Any Other/(μl)Total/(μl)
1111111

1/1

PCR (Target)

 Sukegawa
Templates/(μl)Primer1/(μl)Primer2/(μl)Buffer/(μl)MilliQ/(μl)Any Other/(μl)Total/(μl)
1111111

1/1

PCR (Target)

 Sukegawa
Templates/(μl)Primer1/(μl)Primer2/(μl)Buffer/(μl)MilliQ/(μl)Any Other/(μl)Total/(μl)
1111111

1/1

PCR (Target)

 Sukegawa
Templates/(μl)Primer1/(μl)Primer2/(μl)Buffer/(μl)MilliQ/(μl)Any Other/(μl)Total/(μl)
1111111

1/1

PCR (Target)

 Sukegawa
Templates/(μl)Primer1/(μl)Primer2/(μl)Buffer/(μl)MilliQ/(μl)Any Other/(μl)Total/(μl)
1111111

1/1

PCR (Target)

 Sukegawa
Templates/(μl)Primer1/(μl)Primer2/(μl)Buffer/(μl)MilliQ/(μl)Any Other/(μl)Total/(μl)
1111111

1/1

PCR (Target)

 Sukegawa
Templates/(μl)Primer1/(μl)Primer2/(μl)Buffer/(μl)MilliQ/(μl)Any Other/(μl)Total/(μl)
1111111

1/1

PCR (Target)

 Sukegawa
Templates/(μl)Primer1/(μl)Primer2/(μl)Buffer/(μl)MilliQ/(μl)Any Other/(μl)Total/(μl)
1111111