Difference between revisions of "Team:NCKU Tainan/Notebook Construction"

Revision as of 01:24, 18 October 2016

Notebook Construction - iGEM NCKU

NOTE / Construction

Notebook - Construction

Extracted plasmid from the bacteria provided by XMU
(pSB1C3-BBa_B0034, pSB1C3-BBa_B0015, pSB1C3-BBa_K592009)

Mar. 30/2016

We got the bacteria with Biobrick from XMU.

Apr. 01/2016

Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.

Apr. 02/2016

Culture: 10 ml of LB (with Chloramphenicol) culture with 100 ml/μL pre-cultured bacteria.

And grow up at 37 degree for 10-12 hours.

Apr. 03/2016

Extracted the plasmid, and measured the concentration.

Store the plasmid at -20 degree.

Got the glucose sensitive promoter (PI, Pcar) by PCR

Apr. 16/2016

PCR for PI & Pcar.
Result: After gel purified, the concentration of the DNA was too low to do construction.

Apr. 17/2016

PCR for PI & Pcar again (in big scale).

Purified The PCR product and cloned to T-vector.

Apr. 18/2016

Confirmed the white colony by PCR (using M13F & M13R as the primer)
Result: Some of the colonies were correct.

Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.

Apr. 19/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pMD19T-PI and pMD19T-Pcar were correct.

Apr. 20/2016

Culture: 10 ml of LB (with Ampicillin) cultured with 100 ml/μL pre-cultured bacteria.

Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.

Constructed pSB1C3-B0034-K592009

Apr. 30/2016

Digested pSB1C3-B0034 (SpeI,PstI) and pSB1C3-K592009 (XbaI,PstI)

Ligated K592009 (insert) to pSB1C3-B0034 (vector) and transformed the product.

May 01/2016

Confirmed by colony PCR (using Prefix-F & Suffix-R as the primer).

Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one correct colony from the agar plate.

May 02/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pSB1C3-B0034-K592009 was correct.

May 03/2016

Culture: 10 ml of LB (with Chloramphenicol) cultured with 100ml/μL pre-cultured bacteria.

Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.

Construct pMD19T-Pcar-B0034-K592009 and pMD19T-PI-B0034-K592009

May 07/2016

Digested pSB1C3-B0034-K592009 (XbaI,PstI), pMD19T-Pcar (SpeI,PstI) and pMD19T-PI (SpeI,PstI)

Ligated B0034-K592009 (insert) to pMD19T-Pcar (vector) and pMD19T-Pcar (vector).

Transformed the product.

May 08/2016

Confirmed by colony PCR (using M13F & M13R as the primer).

Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.

May 09/2016

Extracted plasmid and confirm plasmid size by gel electrophoresis.
Result: The size of pMD19T-Pcar-B0034-K592009 and pMD19T-PI-B0034-K592009 were correct.

May 10/2016

Culture: 10 ml of LB (with Ampicillin) cultured with 100 ml/μL pre-cultured bacteria.

Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.

We transformed BioBricks from the 2016 igem distribution
(BBa_J23110, BBa_J23101,BBa_B0031,BBa_B0032,BBa_E1010)

Jun. 03/2016

Transformed 3 ml/μL biobrick to 100 ml/μL DH5α competent cell by heat shock.
Result: The transformation of BBa_J23110, BBa_B0032 and BBa_E1010 was successful.
Result: The transformation of BBa_J23110, BBa_B0032 and BBa_E1010 was fail.

Jun. 04/2016

Pre-culture: 4 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.

Extracted plasmid by Plasmid miniPREP Kit and confirmed plasmid size by gel electrophoresis.

Jun. 05/2016

Culture: 10 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate. And grow up at 37 degree for 10-12 hours.

Jun. 06/2016

Extracted the plasmid, and measure the concentration.

Stored the plasmid at -20 degree.

Jun. 07/2016

Transformed 5 ml/μL biobrick to 100 ml/μL DH5α competent cell by heat shock.
(BBa_J23110, BBa_B0032 and BBa_E1010)
Result: The transformation of BBa_J23110, BBa_B0032 and BBa_E1010 were successful.

Jun. 08/2016

Pre-culture: 4 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.

Extracted plasmid by Plasmid miniPREP Kit and confirmed plasmid size by gel electrophoresis.

Jun. 09/2016

Culture: 10 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.

And grow up at 37 degree for 10-12 hours.

Jun. 10/2016

Extracted the plasmid, and measure the concentration.

Stored the plasmid at -20 degree.

Constructed pSB1C3-B0034-E1010

Jul. 01/2016

Digested pSB1C3-B0034 (SpeI,PstI) and pSB1C3-E1010 (XbaI,PstI)

Ligated E1010 (insert) to pSB1C3-B0034 (vector) and transformed the product.

Jul. 02/2016

Confirmed by colony PCR (using Prefix-F & Suffix-R as the primer).

Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.

Jul. 03/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pSB1C3-B0034-E1010 is correct.

Jul. 04/2016

Culture: 10 ml of LB (with Chloramphenicol) cultured with 100 μl pre-cultured bacteria.

Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.

Extracted plasmid and stored at -20 refrigerator.

Constructed pMD19T-Pcar-B0034-E1010 and pMD19T-PI-B0034-E1010

Jul. 05/2016

Digested pMD19T-Pcar, pMD19T-PI (SpeI,PstI) and pSB1C3-B0034-E1010 (XbaI,PstI)

Ligated B0034-E1010 (insert) to pMD19T-Pcar, pMD19T-PI (vector) and transformed the product.

Jul. 06/2016

Confirmed by colony PCR (using Prefix-F & Suffix-R as the primer).
Result: The length of the PCR product of pMD19T-Pcar-B0034-E1010 was incorrect.

Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one correct colony from the agar plate.

Jul. 07/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pMD19T-PI-B0034-E1010 was correct.

Jul. 08/2016

Culture: 10 ml of LB (with Chloramphenicol) cultured with 100 μl pre-cultured bacteria.

Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.

Extracted plasmid and stored at -20 refrigerator.

Constructed pMD19T-Pcar-B0034-E1010 again

Jul. 11/2016

Digested pMD19T-Pcar (SpeI,PstI) and pSB1C3-B0034-E1010 (XbaI,PstI)

Ligated B0034-E1010 (insert) to pMD19T-Pcar (vector) and transformed the product.

Jul. 12/2016

Confirmed by colony PCR (using Prefix-F & Suffix-R as the primer).

Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one correct colony from the agar plate.

Jul. 13/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pMD19T-Pcar-B0034-E1010 was correct.

Jul. 14/2016

Culture: 10 ml of LB (with Ampicillin) cultured with 100 μl pre-cultured bacteria.

Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.

Extracted plasmid and stored at -20 refrigerator.

Constructed pSB1C3-Pcar-B0034-E1010-TT and pSB1C3-PI-B0034-E1010-TT

Jul. 20/2016

Digested pMD19T-Pcar-B0034-RFP, pMD19T-PI-B0034-RFP (EcoRI, SpeI) and pSB1C3-B0015 (EcoRI, XbaI)

Ligated Pcar-B0034-RFP, PI-B0034-RFP (insert) to pSB1C3-B0015 (vector) and transformed the product.

Jul. 21/2016

Confirmed by colony PCR (using Prefix-F & Suffix-R as the primer).
Result: The PCR result were correct.

Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one correct colony from the agar plate.

Jul. 22/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of two plasmids were correct.

Jul. 23/2016

Culture: 10 ml of LB (with Chloramphenicol) cultured with 100 μl pre-cultured bacteria.

Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.

Extracted plasmid and stored at -20 refrigerator.

We extracted plasmid (pMD19T-P_BAD-lysis-TT) from the bacteria provided by Dr.NG

Jul. 27/2016

Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one colony from LB agar plate.

Jul. 28/2016

Culture: 10 ml of LB (with Ampicillin) was inoculated with one colony from LB agar plate. And grow up at 37 degree for 10-12 hours.

Jul. 29/2016

Extracted the plasmid, and measure the concentration.

Stored the plasmid at -20 degree.

Constructed pSB1A2-B0031-lysis-TT and pSB1A2-B0032-lysis-TT

Aug. 01/2016

PCR for lysis-TT and digested the PCR product (XbaI, PstI).

Digested pSB1A2-B0031 and pSB1A2-B0032 (SpeI, PstI).

Ligated lysis-TT to pSB1A2-B0031 and pSB1A2-B0032 and transformed the product.

Aug. 02/2016

Confirmed by colony PCR (using Prefix-F & Suffix-R as the primer).
Result: The PCR result of pSB1A2-B0031-lysis-TT was correct but pSB1A2-B0032-lysis-TT was fail.

Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.

Aug. 03/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pSB1A2-B0031-lysis-TT were correct.

Aug. 04/2016

Culture: 10 ml of LB (with Chloramphenicol) cultured with 100 μl pre-cultured bacteria.

Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.

Extracted plasmid and stored at -20 refrigerator.

Constructed pSB1A2-B0032-lysis-TT

Aug. 05/2016

PCR for lysis-TT and digested the PCR product (XbaI, PstI).

Digested pSB1A2-B0032 (SpeI, PstI).

Ligated lysis-TT to pSB1A2-B0032 and transformed the product.

Aug. 06/2016

Confirmed by colony PCR (using Prefix-F & Suffix-R as the primer).
Result: The PCR result was correct.

Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.

Aug. 07/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pSB1A2-B0031-lysis-TT were correct.

Aug. 08/2016

Culture: 10 ml of LB (with Chloramphenicol) cultured with 100 μl pre-cultured bacteria.

Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.

Extracted plasmid and stored at -20 refrigerator.

Constructed pSB1C3-J23110-B0031-lysis-TT and pSB1C3-J23110-B0032-lysis-TT
Constructed pSB1C3-J23110-B0031-lysis-TT and pSB1C3-J23110-B0032-lysis-TT

Aug. 09/2016

Digested pSB1A2-B0031-lysis-TT and pSB1A2-B0032-lysis-TT (SpeI, PstI).

Digested pSB1C3-J23101 and pSB1C3-J23110 (XbaI, PstI).

Ligated B0031-lysis-TT and B0032-lysis-TT to pSB1C3-J23101 and transformed the product.

Ligated B0031-lysis-TT and B0032-lysis-TT to pSB1C3-J23110 and transformed the product.

Aug. 10/2016

Confirmed by colony PCR (using Prefix-F & Suffix-R as the primer).
Result: The PCR result of pSB1C3-J23101-B0031-lysis-TT and pSB1C3-J23101-B0032-lysis-TT were correct.
Result: The length of the PCR product of pSB1C3-J23110-B0031-lysis-TT and pSB1C3-J23110-B0032-lysis-TT were incorrect.

Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one correct colony from the agar plate.

Aug. 11/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pSB1C3-J23101-B0031-lysis-TT and pSB1C3-J23101-B0032-lysis-TT were incorrect.

Aug. 12/2016

Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with another one correct colony from the agar plate.

Aug. 13/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pSB1C3-J23101-B0031-lysis-TT and pSB1C3-J23101-B0032-lysis-TT were incorrect.

Constructed pSB1C3-J23101-B0031-lysis-TT and pSB1C3-J23101-B0032-lysis-TT again

Aug. 15/2016

Digested pSB1A2-B0031-lysis-TT and pSB1A2-B0032-lysis-TT (SpeI, PstI).

Digested pSB1C3-J23101 (XbaI, PstI).

Ligated B0031-lysis-TT and B0032-lysis-TT to pSB1C3-J23101 and transformed the product.

Aug. 16/2016

Confirmed by colony PCR (using Prefix-F & Suffix-R as the primer).
Result: The PCR result of pSB1C3-J23101-B0031-lysis-TT and pSB1C3-J23101-B0032-lysis-TT were correct.

Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one correct colony from the agar plate.

Aug. 17/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pSB1C3-J23101-B0031-lysis-TT and pSB1C3-J23101-B0032-lysis-TT were incorrect.

Aug. 18/2016

Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with another one correct colony from the agar plate.

Aug. 19/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pSB1C3-J23101-B0031-lysis-TT and pSB1C3-J23101-B0032-lysis-TT were incorrect.

Constructed pMD19T-Pcar-E1010-B0015-P_BAD-lysis-TT (reverse) and
pMD19T-PI-E1010-B0015-P_BAD-lysis-TT (reverse)

Aug. 24/2016

PCR for Pcar-E1010-B0015 and PI-E1010-B0015.

Digested the PCR product (EcoRI, XbaI).

Digested pMD19T -P_BAD-lysis-TT (EcoRI, XbaI).

Ligated Pcar-E1010-B0015 and PI-E1010-B0015 to pMD19T -P_BAD-lysis-TT and transformed the product.

Aug. 25/2016

Confirmed by colony PCR (using Prefix-F & Suffix-R as the primer).
Result: The PCR results were correct.

Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.

Aug. 26/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pMD19T-Pcar-E1010-B0015-P_BAD-lysis-TT and pMD19T-PI-E1010-B0015-P_BAD-lysis-TT were correct.

Aug. 27/2016

Culture: 10 ml of LB (with Ampicillin) cultured with 100 μl pre-cultured bacteria.

Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.

Extracted plasmid and stored at -20 refrigerator.

Aug. 28/2016

Transformed plasmid (pMD19T-Pcar-E1010-B0015-P_BAD-lysis-TT and pMD19T-PI-E1010-B0015-P_BAD-lysis-TT) to BL21 competent cell.

Aug. 29/2016

Pre-culture: 4 ml of LB (with Ampicillin) was inoculated with one colony from the agar plate.

Aug. 30/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pMD19T-Pcar-E1010-B0015-P_BAD-lysis-TT and pMD19T-PI-E1010-B0015-P_BAD-lysis-TT were correct.

Aug. 31/2016

Culture: 10 ml of LB (with Ampicillin) cultured with 100 μl pre-cultured bacteria.

Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.

Extracted plasmid and stored at -20 refrigerator.

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+     <style>@font-face {  font-family: 'NotoSansCJKtc-Regular';  src: url("/wiki/images/0/0b/T--NCKU_Tainan--NotoSansCJKtc-Regular.woff") format('woff');}</style><nav class="navbar navbar-default"><div class="container-fluid" style="width:72%">  <div class="navbar-header">      <button type="button" class="navbar-toggle collapsed" data-toggle="collapse" data-target="#navbar" aria-expanded="false">        <span class="sr-only">Toggle navigation</span>        <span class="icon-bar"></span>        <span class="icon-bar"></span>        <span class="icon-bar"></span>      </button>      <a class="navbar-brand" href="/Team:NCKU_Tainan">        <h1>NCKU</h1><h4>Tainan</h4>      </a>   </div>  <div id="navbar" class="navbar-collapse collapse">    <ul class="nav navbar-nav">      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu1" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Project</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu2">          <li><a href="/Team:NCKU_Tainan/Project">Background</a></li>          <li><a href="/Team:NCKU_Tainan/Description">Description</a></li>          <li><a href="/Team:NCKU_Tainan/Results">Results</a></li>          <li><a href="/Team:NCKU_Tainan/Model">Modeling</a></li>          <li><a href="/Team:NCKU_Tainan/Parts">Parts</a></li>        </ul>      </li>      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu2" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Device</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu2">          <li><a href="/Team:NCKU_Tainan/Hardware">Hardware</a></li>          <li><a href="/Team:NCKU_Tainan/Software">Software</a></li>          <li><a href="/Team:NCKU_Tainan/Demonstrate">Demonstrate</a></li>        </ul>      </li>       <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu3" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Judging</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu3">          <li><a href="/Team:NCKU_Tainan/Medal">Medal</a></li>          <li><a href="/Team:NCKU_Tainan/Safety">Safety</a></li>        </ul>        </li>      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu4" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="/Team:NCKU_Tainan/Team">Team</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu4">          <li><a href="/Team:NCKU_Tainan/Team">Team</a></li>          <li><a href="/Team:NCKU_Tainan/Attributions">Attributions</a></li>          <li><a href="/Team:NCKU_Tainan/Acknowledgement">Acknowledgement</a></li>          <li><a href="/Team:NCKU_Tainan/Collaborations">Collaborations</a></li>        </ul>        </li>      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu5" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="/Team:NCKU_Tainan/Human_Practices">Human Practices</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu5">          <li><a href="/Team:NCKU_Tainan/Human_Practices">Overview</a></li>          <li><a href="/Team:NCKU_Tainan/Integrated_Practices">Integrated Practices</a></li>          <li><a href="/Team:NCKU_Tainan/Engagement">Engagement</a></li>        </ul>      </li>      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu6" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Notebook</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu6">          <li><a href="/Team:NCKU_Tainan/Notebook_Construction">Construction</a></li>          <li><a href="/Team:NCKU_Tainan/Notebook_Functional_Test">Functional Test</a></li>          <li><a href="/Team:NCKU_Tainan/Notebook_Device_Design">Device Design</a></li>          <li><a href="/Team:NCKU_Tainan/Notebook_Protocols">Protocols</a></li>          <li><a href="/Team:NCKU_Tainan/Notebook_Model">Model</a></li>        </ul>      </li>    </ul>    <ul class="nav navbar-nav navbar-right">    </ul>  </div><!--/.nav-collapse --></div><!--/.container-fluid --></nav><div id="container-big"><div id="iGEMbar"></div><div id="line-left"></div><div id="line-left2"></div><div id="line-right"></div><div id="photo-left"></div></div><!--/.container-big -->        <style>
-			#photo-left { background-image: url("/wiki/images/c/c1/T--NCKU_Tainan--sample2.jpg"); }
+            #photo-left { background-image: url("/wiki/images/0/0c/T--NCKU_Tainan--Construction.png"); }
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-		<div class="container-fluid" style="margin-top:100px">
+        <div class="container-fluid" style="margin-top:100px">
-				<div class="head">NOTE / Construction</div>
+                <div class="head">NOTE / Construction</div>
-				<div class="content row">
+                <div class="content row">
-					<div class="col-md-9">
+                    <div class="col-md-9">
-						<div class="head2">Notebook - Construction</div>
+                        <div class="head2">Notebook - Construction</div>
-						<div class="title-line" id="sec1"></div>
+                        <div class="title-line long" id="sec1"></div>
-						<div class="title-content">Extracted plasmid from the bacteria provided by XMU<br>(pSB1C3-BBa_B0034, pSB1C3-BBa_B0015, pSB1C3-BBa_K592009)</div>
+                        <div class="title-content">Extracted plasmid from the bacteria provided by XMU<br>(pSB1C3-BBa_B0034, pSB1C3-BBa_B0015, pSB1C3-BBa_K592009)</div>
-						<h5>2016.03.30</h5>
+												<div class="text-content">
-						<p>We got the bacteria with Biobrick from XMU.</p>
+                          <h5>Mar. 30/2016</h5>
-						<h5>2016.04.01</h5>
+                          <p>We got the bacteria with Biobrick from XMU.</p>
-						<p>Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.</p>
+                          <h5>Apr. 01/2016</h5>
-						<h5>2016.04.02</h5>
+                          <p>Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.</p>
-						<p>Culture: 10 ml of LB (with Chloramphenicol) culture with 100 ml/μL pre-cultured bacteria. </p>
+                          <h5>Apr. 02/2016</h5>
-						<p>And grow up at 37 degree for 10-12 hours.</p>
+                          <p>Culture: 10 ml of LB (with Chloramphenicol) culture with 100 ml/μL pre-cultured bacteria. </p>
-						<h5>2016.04.03</h5>
+                          <p>And grow up at 37 degree for 10-12 hours.</p>
-						<p>Extracted the plasmid, and measured the concentration.</p>
+                          <h5>Apr. 03/2016</h5>
-						<p>Store the plasmid at -20 degree.</p>
+                          <p>Extracted the plasmid, and measured the concentration.</p>
+                          <p>Store the plasmid at -20 degree.</p>
-						<div class="title-line" id="sec2"></div>
+                        </div>
-						<div class="title-content">Got the glucose sensitive promoter (PpI, Pcar) by PCR</div>
+                        <div class="title-line long" id="sec2"></div>
-						<h5>2016.04.16</h5>
+                        <div class="title-content">Got the glucose sensitive promoter (PI, Pcar) by PCR</div>
-						<p>PCR for PpI &amp; Pcar.<br>
+												<div class="text-content">
+                          <h5>Apr. 16/2016</h5>
+                          <p>PCR for PI &amp; Pcar.<br>
                 <i class="glyphicon glyphicon-ok"></i>Result: After gel purified, the concentration of the DNA was too low to do construction.</p>
-						<h5>2016.04.17</h5>
+                          <h5>Apr. 17/2016</h5>
-						<p>PCR for PpI &amp; Pcar again (in big scale).</p>
+                          <p>PCR for PI &amp; Pcar again (in big scale).</p>
-             <p>Purified the PCR product and cloned to T-vector.</p>
+             <p>Purified   The PCR product and cloned to T-vector.</p>
-						<h5>2016.04.18</h5>
+                          <h5>Apr. 18/2016</h5>
-						<p>Confirmed the white colony by PCR (using M13F &amp; M13R as the primer)<br>
+                          <p>Confirmed the white colony by PCR (using M13F &amp; M13R as the primer)<br>
-               <i class="glyphicon glyphicon-ok"></i>Result: Some of the colonies were correct.</p>
+                            <i class="glyphicon glyphicon-ok"></i>Result: Some of the colonies were correct.</p>
              <p>Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.</p>
-						<h5>2016.04.19</h5>
+                          <h5>Apr. 19/2016</h5>
-						<p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
+                          <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
-               <i class="glyphicon glyphicon-ok"></i>Result: The size of pMD19T-PpI and pMD19T-Pcar were correct.</p>
+                            <i class="glyphicon glyphicon-ok"></i>Result: The size of pMD19T-PI and pMD19T-Pcar were correct.</p>
-						<h5>2016.04.20</h5>
+                          <h5>Apr. 20/2016</h5>
-						<p>Culture: 10 ml of LB (with Ampicillin) cultured with 100 ml/μL pre-cultured bacteria.</p>
+                          <p>Culture: 10 ml of LB (with Ampicillin) cultured with 100 ml/μL pre-cultured bacteria.</p>
-            <p>Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.</p>
+                          <p>Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.</p>
-            <div class="title-line" id="sec3"></div>
+												</div>
-						<div class="title-content">Constructed pSB1C3-B0034-K592009</div>
-						<h5>2016.04.30</h5>
+                        <div class="title-line long" id="sec3"></div>
-            <p>Digested pSB1C3-B0034 (<i>SpeI,PstI</i>) and pSB1C3-K592009 (<i>XbaI,PstI</i>)</p>
+                        <div class="title-content">Constructed pSB1C3-B0034-K592009</div>
-            <p>Ligated K592009 (insert) to pSB1C3-B0034 (vector) and transformed the product.</p>
+												<div class="text-content">
-            <h5>2016.05.01</h5>
+                          <h5>Apr. 30/2016</h5>
+                          <p>Digested pSB1C3-B0034 (<i>SpeI,PstI</i>) and pSB1C3-K592009 (<i>XbaI,PstI</i>)</p>
+                          <p>Ligated K592009 (insert) to pSB1C3-B0034 (vector) and transformed the product.</p>
+                          <h5>May 01/2016</h5>
+                          <p>Confirmed by colony PCR (using Prefix-F &amp; Suffix-R as the primer).</p>
+                          <p>Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one correct colony from the agar plate.</p>
+                          <h5>May 02/2016</h5>
+                          <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
+                             <i class="glyphicon glyphicon-ok"></i>Result: The size of pSB1C3-B0034-K592009 was correct.
+</p>
+                          <h5>May 03/2016</h5>
+                          <p>Culture: 10 ml of LB (with Chloramphenicol) cultured with 100ml/μL pre-cultured bacteria.</p>
+                          <p>Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.</p>
+                        </div>
+                        <div class="title-line long" id="sec4"></div>
+                        <div class="title-content">Construct pMD19T-Pcar-B0034-K592009 and pMD19T-PI-B0034-K592009</div>
+                        <div class="text-content">
+                          <h5>May 07/2016</h5>
+                          <p>Digested pSB1C3-B0034-K592009 (<i>XbaI,PstI</i>), pMD19T-Pcar (<i>SpeI,PstI</i>) and pMD19T-PI (<i>SpeI,PstI</i>)</p>
+                          <p>Ligated B0034-K592009 (insert) to pMD19T-Pcar (vector) and pMD19T-Pcar (vector). </p>
+                          <p>Transformed the product.</p>
+                          <h5>May 08/2016</h5>
+                          <p>Confirmed by colony PCR (using M13F &amp; M13R as the primer).</p>
+                          <p>Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.</p>
+                          <h5>May 09/2016</h5>
+                          <p>Extracted plasmid and confirm plasmid size by gel electrophoresis.<br>
+                             <i class="glyphicon glyphicon-ok"></i>Result: The size of pMD19T-Pcar-B0034-K592009 and pMD19T-PI-B0034-K592009 were correct.</p>
+                          <h5>May 10/2016</h5>
+                          <p>Culture: 10 ml of LB (with Ampicillin) cultured with 100 ml/μL pre-cultured bacteria.</p>
+                          <p>Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.</p>
+                        </div>
+                        <div class="title-line long" id="sec5"></div>
+                        <div class="title-content">We transformed BioBricks from the 2016 igem distribution<br>
+              (BBa_J23110, BBa_J23101,BBa_B0031,BBa_B0032,BBa_E1010)</div>
+                        <div class="text-content">
+                          <h5>Jun. 03/2016</h5>
+                          <p>Transformed 3 ml/μL biobrick to 100 ml/μL DH5α competent cell by heat shock.<br>
+                             <i class="glyphicon glyphicon-ok"></i>Result: The transformation of BBa_J23110, BBa_B0032 and BBa_E1010 was successful.<br>
+                             <i class="glyphicon glyphicon-remove"></i>Result: The transformation of BBa_J23110, BBa_B0032 and BBa_E1010 was fail.</p>
+                          <h5>Jun. 04/2016</h5>
+                          <p>Pre-culture: 4 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.</p>
+                          <p>Extracted plasmid by Plasmid miniPREP Kit and confirmed plasmid size by gel electrophoresis.</p>
+                          <h5>Jun. 05/2016</h5>
+                          <p>Culture: 10 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate. And grow up at 37 degree for 10-12 hours.</p>
+                          <h5>Jun. 06/2016</h5>
+                          <p>Extracted the plasmid, and measure the concentration.</p>
+                          <p>Stored the plasmid at -20 degree.</p>
+                          <h5>Jun. 07/2016</h5>
+                          <p>Transformed 5 ml/μL biobrick to 100 ml/μL DH5α competent cell by heat shock.<br>
+                            (BBa_J23110, BBa_B0032 and BBa_E1010)<br>
+                            <i class="glyphicon glyphicon-ok"></i>Result: The transformation of BBa_J23110, BBa_B0032 and BBa_E1010 were successful.</p>
+                          <h5>Jun. 08/2016</h5>
+                          <p>Pre-culture: 4 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.</p>
+                          <p>Extracted plasmid by Plasmid miniPREP Kit and confirmed plasmid size by gel electrophoresis.</p>
+                          <h5>Jun. 09/2016</h5>
+                          <p>Culture: 10 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.</p>
+                          <p>And grow up at 37 degree for 10-12 hours.</p>
+                          <h5>Jun. 10/2016</h5>
+                          <p>Extracted the plasmid, and measure the concentration.</p>
+                          <p>Stored the plasmid at -20 degree.</p>
+                        </div>
+            <div class="title-line long" id="sec6"></div>
+            <div class="title-content">Constructed pSB1C3-B0034-E1010</div>
+                        <div class="text-content">
+            <h5>Jul. 01/2016</h5>
+            <p>Digested pSB1C3-B0034 (<i>SpeI,PstI</i>) and pSB1C3-E1010 (<i>XbaI,PstI</i>)</p>
+            <p>Ligated E1010 (insert) to pSB1C3-B0034 (vector) and transformed the product.</p>
+            <h5>Jul. 02/2016</h5>
              <p>Confirmed by colony PCR (using Prefix-F &amp; Suffix-R as the primer).</p>
+            <p>Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.</p>
+            <h5>Jul. 03/2016</h5>
+            <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
+            <i class="glyphicon glyphicon-ok"></i>Result: The size of pSB1C3-B0034-E1010 is correct.</p>
+            <h5>Jul. 04/2016</h5>
+            <p>Culture: 10 ml of LB (with Chloramphenicol) cultured with 100 μl pre-cultured bacteria.</p>
+            <p>Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator. </p>
+            <p>Extracted plasmid and stored at -20 refrigerator.</p>
+                        </div>
+            <div class="title-line long" id="sec7"></div>
+            <div class="title-content">Constructed pMD19T-Pcar-B0034-E1010 and pMD19T-PI-B0034-E1010</div>
+                        <div class="text-content">
+            <h5>Jul. 05/2016</h5>
+            <p>Digested pMD19T-Pcar, pMD19T-PI (SpeI,PstI) and pSB1C3-B0034-E1010 (<i>XbaI,PstI</i>)</p>
+            <p>Ligated B0034-E1010 (insert) to pMD19T-Pcar, pMD19T-PI (vector) and transformed the product.</p>
+            <h5>Jul. 06/2016</h5>
+            <p>Confirmed by colony PCR (using Prefix-F &amp; Suffix-R as the primer).<br>
+            <i class="glyphicon glyphicon-remove"></i>Result: The length of the PCR product of pMD19T-Pcar-B0034-E1010 was incorrect.</p>
              <p>Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one correct colony from the agar plate.</p>
-             <h5>2016.05.02</h5>
+             <h5>Jul. 07/2016</h5>
              <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
-               <i class="glyphicon glyphicon-ok"></i>Result: The size of pSB1C3-B0034-K592009 was correct.
+            <i class="glyphicon glyphicon-ok"></i>Result: The size of pMD19T-PI-B0034-E1010 was correct.</p>
-</p>
+             <h5>Jul. 08/2016</h5>
-             <h5>2016.05.03</h5>
+             <p>Culture: 10 ml of LB (with Chloramphenicol) cultured with 100 μl pre-cultured bacteria.</p>
-             <p>Culture: 10 ml of LB (with Chloramphenicol) cultured with 100ml/μL pre-cultured bacteria.</p>
              <p>Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.</p>
+            <p>Extracted plasmid and stored at -20 refrigerator.</p>
+                        </div>
-             <div class="title-line" id="sec3"></div>
+             <div class="title-line long" id="sec8"></div>
-						<div class="title-content">Construct pMD19T-Pcar-B0034-K592009 and pMD19T-PpI-B0034-K592009</div>
+            <div class="title-content">Constructed pMD19T-Pcar-B0034-E1010 again</div>
-						<h5>2016.05.07</h5>
+                        <div class="text-content">
-             <p>Digested pSB1C3-B0034-K592009 (<i>XbaI,PstI</i>), pMD19T-Pcar (<i>SpeI,PstI</i>) and pMD19T-PpI (<i>SpeI,PstI</i>)
+            <h5>Jul. 11/2016</h5>
-            </p><p>Ligated B0034-K592009 (insert) to pMD19T-Pcar (vector) and pMD19T-Pcar (vector). </p>
+             <p>Digested pMD19T-Pcar (<i>SpeI,PstI</i>) and pSB1C3-B0034-E1010 (<i>XbaI,PstI</i>)</p>
-            <p>Transformed the product.</p>
+            <p>Ligated B0034-E1010 (insert) to pMD19T-Pcar (vector) and transformed the product.</p>
-             <h5>2016.05.08</h5>
+             <h5>Jul. 12/2016</h5>
-             <p>Confirmed by colony PCR (using M13F &amp; M13R as the primer).</p>
+             <p>Confirmed by colony PCR (using Prefix-F &amp; Suffix-R as the primer).</p>
-             <p>Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.</p>
+             <p>Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one correct colony from the agar plate.</p>
-             <h5>2016.05.09</h5>
+             <h5>Jul. 13/2016</h5>
-             <p>Extracted plasmid and confirm plasmid size by gel electrophoresis.<br>
+             <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
-               <i class="glyphicon glyphicon-ok"></i>Result: The size of pMD19T-Pcar-B0034-K592009 and pMD19T-PpI-B0034-K592009 were correct.</p>
+            <i class="glyphicon glyphicon-ok"></i>Result: The size of pMD19T-Pcar-B0034-E1010 was correct.</p>
-             <h5>2016.05.10</h5>
+             <h5>Jul. 14/2016</h5>
-             <p>Culture: 10 ml of LB (with Ampicillin) cultured with 100 ml/μL pre-cultured bacteria.</p>
+             <p>Culture: 10 ml of LB (with Ampicillin) cultured with 100 μl pre-cultured bacteria.</p>
-             <p>Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.</p>
+             <p>Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator. </p>
+            <p>Extracted plasmid and stored at -20 refrigerator.</p>
+                        </div>
-             <div class="title-line" id="sec3"></div>
+             <div class="title-line long" id="sec9"></div>
-						<div class="title-content">We transformed BioBricks from the 2016 igem distribution<br>
+            <div class="title-content">Constructed pSB1C3-Pcar-B0034-E1010-TT and pSB1C3-PI-B0034-E1010-TT</div>
-              (BBa_J23110, BBa_J23101,BBa_B0031,BBa_B0032,BBa_E1010)</div>
+                        <div class="text-content">
-             <h5>2016.06.03</h5>
+             <h5>Jul. 20/2016</h5>
-             <p>Transformed 3 ml/μL biobrick to 100 ml/μL DH5α competent cell by heat shock.<br>
+             <p>Digested pMD19T-Pcar-B0034-RFP, pMD19T-PI-B0034-RFP (<i>EcoRI, SpeI</i>) and pSB1C3-B0015 (<i>EcoRI, XbaI</i>)</p>
-               <i class="glyphicon glyphicon-ok"></i>Result: The transformation of BBa_J23110, BBa_B0032 and BBa_E1010 was successful.<br>
+            <p>Ligated Pcar-B0034-RFP, PI-B0034-RFP (insert) to pSB1C3-B0015 (vector) and transformed the product.</p>
-               <i class="glyphicon glyphicon-remove"></i>Result: The transformation of BBa_J23110, BBa_B0032 and BBa_E1010 was fail.</p>
+            <h5>Jul. 21/2016</h5>
-            <h5>2016.06.04</h5>
+            <p>Confirmed by colony PCR (using Prefix-F &amp; Suffix-R as the primer).<br>
-             <p>Pre-culture: 4 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.</p>
+            <i class="glyphicon glyphicon-ok"></i>Result: The PCR result were correct.</p>
-             <p>Extracted plasmid by Plasmid miniPREP Kit and confirmed plasmid size by gel electrophoresis.</p>
+             <p>Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one correct colony from the agar plate.</p>
-             <h5>2016.06.05</h5>
+            <h5>Jul. 22/2016</h5>
-             <p>Culture: 10 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate. And grow up at 37 degree for 10-12 hours.</p>
+             <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
-            <h5>2016.06.06</h5>
+            <i class="glyphicon glyphicon-ok"></i>Result: The size of two plasmids were correct.</p>
-             <p>Extracted the plasmid, and measure the concentration.</p>
+             <h5>Jul. 23/2016</h5>
-             <p>Stored the plasmid at -20 degree.</p>
+             <p>Culture: 10 ml of LB (with Chloramphenicol) cultured with 100 μl pre-cultured bacteria.</p>
-            <h5>2016.06.07</h5>
+             <p>Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.</p>
-             <p>Transformed 5 ml/μL biobrick to 100 ml/μL DH5α competent cell by heat shock.<br>
+             <p>Extracted plasmid and stored at -20 refrigerator.</p>
-              (BBa_J23110, BBa_B0032 and BBa_E1010)<br>
+                        </div>
-              <i class="glyphicon glyphicon-ok"></i>Result: The transformation of BBa_J23110, BBa_B0032 and BBa_E1010 were successful.</p>
-             <h5>2016.06.08</h5>
+             <div class="title-line long" id="sec10"></div>
-             <p>Pre-culture: 4 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.</p>
+            <div class="title-content">We extracted plasmid (pMD19T-P<sub>BAD</sub>-lysis-TT) from the bacteria provided by Dr.NG</div>
-             <p>Extracted plasmid by Plasmid miniPREP Kit and confirmed plasmid size by gel electrophoresis.</p>
+                        <div class="text-content">
-            <h5>2016.06.09</h5>
+             <h5>Jul. 27/2016</h5>
-             <p>Culture: 10 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.</p>
+             <p>Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one colony from LB agar plate.</p>
-            <p>And grow up at 37 degree for 10-12 hours.</p>
+             <h5>Jul. 28/2016</h5>
-             <h5>2016.06.10</h5>
+             <p>Culture: 10 ml of LB (with Ampicillin) was inoculated with one colony from LB agar plate. And grow up at 37 degree for 10-12 hours.</p>
+             <h5>Jul. 29/2016</h5>
              <p>Extracted the plasmid, and measure the concentration.</p>
              <p>Stored the plasmid at -20 degree.</p>
+                        </div>
+            <div class="title-line long" id="sec11"></div>
+            <div class="title-content">Constructed pSB1A2-B0031-lysis-TT and pSB1A2-B0032-lysis-TT</div>
+                        <div class="text-content">
+            <h5>Aug. 01/2016</h5>
+            <p>PCR for lysis-TT and digested the PCR product (<i>XbaI, PstI</i>). </p>
+            <p>Digested pSB1A2-B0031 and pSB1A2-B0032 (<i>SpeI, PstI</i>).</p>
+            <p>Ligated lysis-TT to pSB1A2-B0031 and pSB1A2-B0032 and transformed the product.</p>
+            <h5>Aug. 02/2016</h5>
+            <p>Confirmed by colony PCR (using Prefix-F &amp; Suffix-R as the primer).<br>
+            <i class="glyphicon glyphicon-ok"></i>Result: The PCR result of pSB1A2-B0031-lysis-TT was correct but pSB1A2-B0032-lysis-TT was fail.</p>
+            <p>Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.</p>
+            <h5>Aug. 03/2016</h5>
+            <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
+            <i class="glyphicon glyphicon-ok"></i>Result: The size of pSB1A2-B0031-lysis-TT were correct.</p>
+            <h5>Aug. 04/2016</h5>
+            <p>Culture: 10 ml of LB (with Chloramphenicol) cultured with 100 μl pre-cultured bacteria.</p>
+            <p>Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator. </p>
+            <p>Extracted plasmid and stored at -20 refrigerator.</p>
+                        </div>
+            <div class="title-line long" id="sec12"></div>
+            <div class="title-content">Constructed pSB1A2-B0032-lysis-TT</div>
+                        <div class="text-content">
+            <h5>Aug. 05/2016</h5>
+            <p>PCR for lysis-TT and digested the PCR product (<i>XbaI, PstI</i>). </p>
+            <p>Digested pSB1A2-B0032 (<i>SpeI, PstI</i>).</p>
+            <p>Ligated lysis-TT to pSB1A2-B0032 and transformed the product.</p>
+            <h5>Aug. 06/2016</h5>
+            <p>Confirmed by colony PCR (using Prefix-F &amp; Suffix-R as the primer).<br>
+            <i class="glyphicon glyphicon-ok"></i>Result: The PCR result was correct.</p>
+            <p>Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.</p>
+            <h5>Aug. 07/2016</h5>
+            <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
+            <i class="glyphicon glyphicon-ok"></i>Result: The size of pSB1A2-B0031-lysis-TT were correct.</p>
+            <h5>Aug. 08/2016</h5>
+            <p>Culture: 10 ml of LB (with Chloramphenicol) cultured with 100 μl pre-cultured bacteria.</p>
+            <p>Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator. </p>
+            <p>Extracted plasmid and stored at -20 refrigerator.</p>
+                        </div>
+            <div class="title-line long" id="sec13"></div>
+            <div class="title-content">Constructed pSB1C3-J23110-B0031-lysis-TT and pSB1C3-J23110-B0032-lysis-TT<br>
+            Constructed pSB1C3-J23110-B0031-lysis-TT and pSB1C3-J23110-B0032-lysis-TT</div>
+                        <div class="text-content">
+            <h5>Aug. 09/2016</h5>
+            <p>Digested pSB1A2-B0031-lysis-TT and pSB1A2-B0032-lysis-TT (<i>SpeI, PstI</i>).</p>
+            <p>Digested pSB1C3-J23101 and pSB1C3-J23110 (<i>XbaI, PstI</i>).</p>
+            <p>Ligated B0031-lysis-TT and B0032-lysis-TT to pSB1C3-J23101 and transformed the product.</p>
+            <p>Ligated B0031-lysis-TT and B0032-lysis-TT to pSB1C3-J23110 and transformed the product.</p>
+            <h5>Aug. 10/2016</h5>
+            <p>Confirmed by colony PCR (using Prefix-F &amp; Suffix-R as the primer).<br>
+            <i class="glyphicon glyphicon-ok"></i>Result: The PCR result of pSB1C3-J23101-B0031-lysis-TT and pSB1C3-J23101-B0032-lysis-TT were correct.<br>
+            <i class="glyphicon glyphicon-reomve"></i>Result: The length of the PCR product of pSB1C3-J23110-B0031-lysis-TT and pSB1C3-J23110-B0032-lysis-TT were incorrect.</p>
+            <p>Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one correct colony from the agar plate.</p>
+            <h5>Aug. 11/2016</h5>
+            <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
+            <i class="glyphicon glyphicon-reomve"></i>Result: The size of pSB1C3-J23101-B0031-lysis-TT and pSB1C3-J23101-B0032-lysis-TT were incorrect.</p>
+            <h5>Aug. 12/2016</h5>
+            <p>Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with another one correct colony from the agar plate.</p>
+            <h5>Aug. 13/2016</h5>
+            <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
+            <i class="glyphicon glyphicon-reomve"></i>Result: The size of pSB1C3-J23101-B0031-lysis-TT and pSB1C3-J23101-B0032-lysis-TT were incorrect.</p>
+                        </div>
+            <div class="title-line long" id="sec14"></div>
+            <div class="title-content">Constructed pSB1C3-J23101-B0031-lysis-TT and pSB1C3-J23101-B0032-lysis-TT again</div>
+                        <div class="text-content">
+            <h5>Aug. 15/2016</h5>
+            <p>Digested pSB1A2-B0031-lysis-TT and pSB1A2-B0032-lysis-TT (<i>SpeI, PstI</i>).</p>
+            <p>Digested pSB1C3-J23101 (<i>XbaI, PstI</i>).</p>
+            <p>Ligated B0031-lysis-TT and B0032-lysis-TT to pSB1C3-J23101 and transformed the product.</p>
+            <h5>Aug. 16/2016</h5>
+            <p>Confirmed by colony PCR (using Prefix-F &amp; Suffix-R as the primer).<br>
+            <i class="glyphicon glyphicon-ok"></i>Result: The PCR result of pSB1C3-J23101-B0031-lysis-TT and pSB1C3-J23101-B0032-lysis-TT were correct.</p>
+            <p>Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one correct colony from the agar plate.</p>
+            <h5>Aug. 17/2016</h5>
+            <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
+            <i class="glyphicon glyphicon-remove"></i>Result: The size of pSB1C3-J23101-B0031-lysis-TT and pSB1C3-J23101-B0032-lysis-TT were incorrect.</p>
+            <h5>Aug. 18/2016</h5>
+            <p>Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with another one correct colony from the agar plate.</p>
+            <h5>Aug. 19/2016</h5>
+            <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
+            <i class="glyphicon glyphicon-remove"></i>Result: The size of pSB1C3-J23101-B0031-lysis-TT and pSB1C3-J23101-B0032-lysis-TT were incorrect.</p>
+                        </div>
+            <div class="title-line long" id="sec15"></div>
+            <div class="title-content">Constructed pMD19T-Pcar-E1010-B0015-P<sub>BAD</sub>-lysis-TT (reverse) and <br>pMD19T-PI-E1010-B0015-P<sub>BAD</sub>-lysis-TT (reverse)</div>
+                        <div class="text-content">
+            <h5>Aug. 24/2016</h5>
+            <p>PCR for Pcar-E1010-B0015 and PI-E1010-B0015.</p>
+            <p>Digested the PCR product (<i>EcoRI, XbaI</i>). </p>
+            <p>Digested pMD19T -P<sub>BAD</sub>-lysis-TT (<i>EcoRI, XbaI</i>).</p>
+            <p>Ligated Pcar-E1010-B0015 and PI-E1010-B0015 to pMD19T -P<sub>BAD</sub>-lysis-TT and transformed the product.</p>
+            <h5>Aug. 25/2016</h5>
+            <p>Confirmed by colony PCR (using Prefix-F &amp; Suffix-R as the primer).<br>
+            <i class="glyphicon glyphicon-ok"></i>Result: The PCR results were correct.</p>
+            <p>Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.</p>
+            <h5>Aug. 26/2016</h5>
+            <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
+            <i class="glyphicon glyphicon-ok"></i>Result: The size of pMD19T-Pcar-E1010-B0015-P<sub>BAD</sub>-lysis-TT and pMD19T-PI-E1010-B0015-P<sub>BAD</sub>-lysis-TT were correct.</p>
+            <h5>Aug. 27/2016</h5>
+            <p>Culture: 10 ml of LB (with Ampicillin) cultured with 100 μl pre-cultured bacteria.</p>
+            <p>Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator. </p>
+            <p>Extracted plasmid and stored at -20 refrigerator.</p>
+            <h5>Aug. 28/2016</h5>
+            <p>Transformed plasmid (pMD19T-Pcar-E1010-B0015-P<sub>BAD</sub>-lysis-TT and pMD19T-PI-E1010-B0015-P<sub>BAD</sub>-lysis-TT) to BL21 competent cell.</p>
+            <h5>Aug. 29/2016</h5>
+            <p>Pre-culture: 4 ml of LB (with Ampicillin) was inoculated with one colony from the agar plate.</p>
+            <h5>Aug. 30/2016</h5>
+            <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
+            <i class="glyphicon glyphicon-ok"></i>Result: The size of pMD19T-Pcar-E1010-B0015-P<sub>BAD</sub>-lysis-TT and pMD19T-PI-E1010-B0015-P<sub>BAD</sub>-lysis-TT were correct.</p>
+            <h5>Aug. 31/2016</h5>
+            <p>Culture: 10 ml of LB (with Ampicillin) cultured with 100 μl pre-cultured bacteria.</p>
+            <p>Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator. </p>
+            <p>Extracted plasmid and stored at -20 refrigerator.</p>
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