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

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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>
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<div class="head">NOTEBOOK / Overview</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>
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<div class="head2">Notebook - Overview</div>
<div class="title-line" id="sec1"></div>
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<div class="intro">We divide our record - notebook into four parts:</div>
<div class="title-content">Extracted plasmid from the bacteria provided by XMU<br>(pSB1C3-BBa_B0034, pSB1C3-BBa_B0015, pSB1C3-BBa_K592009)</div>
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<ol>
<h5>2016.03.30</h5>
+
<li><a href="/Team:NCKU_Tainan/Notebook_Construction">Construction</a></li>
<p>We got the bacteria with Biobrick from XMU.</p>
+
<li><a href="/Team:NCKU_Tainan/Notebook_Functional_Test">Functional Test</a></li>
<h5>2016.04.01</h5>
+
<li><a href="/Team:NCKU_Tainan/Notebook_Device_Design">Device Design</a></li>
<p>Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.</p>
+
<li><a href="#">Protocols</a></li>
<h5>2016.04.02</h5>
+
</ol>
<p>Culture: 10 ml of LB (with Chloramphenicol) culture with 100 ml/μL pre-cultured bacteria. </p>
+
<p>And grow up at 37 degree for 10-12 hours.</p>
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<h5>2016.04.03</h5>
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<p>Extracted the plasmid, and measured the concentration.</p>
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<p>Store the plasmid at -20 degree.</p>
+
 
+
<div class="title-line" id="sec2"></div>
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<div class="title-content">Got the glucose sensitive promoter (PpI, Pcar) by PCR</div>
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<h5>2016.04.16</h5>
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<p>PCR for PpI &amp; Pcar.<br>
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              <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>
+
<p>PCR for PpI &amp; Pcar again (in big scale).</p>
+
            <p>Purified the PCR product and cloned to T-vector.</p>
+
<h5>2016.04.18</h5>
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<p>Confirmed the white colony by PCR (using M13F &amp; M13R as the primer)<br>
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              <i class="glyphicon glyphicon-ok"></i>Result: Some of the colonies were correct.</p>
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            <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>
+
<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>
+
<h5>2016.04.20</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 class="title-line" id="sec3"></div>
+
<div class="title-content">Constructed pSB1C3-B0034-K592009</div>
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<h5>2016.04.30</h5>
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            <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>2016.05.01</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>2016.05.02</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>2016.05.03</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 class="title-line" id="sec3"></div>
+
<div class="title-content">Construct pMD19T-Pcar-B0034-K592009 and pMD19T-PpI-B0034-K592009</div>
+
<h5>2016.05.07</h5>
+
            <p>Digested pSB1C3-B0034-K592009 (<i>XbaI,PstI</i>), pMD19T-Pcar (<i>SpeI,PstI</i>) and pMD19T-PpI (<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>2016.05.08</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>2016.05.09</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-PpI-B0034-K592009 were correct.</p>
+
            <h5>2016.05.10</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 class="title-line" id="sec3"></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>
+
            <h5>2016.06.03</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>2016.06.04</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>2016.06.05</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>2016.06.06</h5>
+
            <p>Extracted the plasmid, and measure the concentration.</p>
+
            <p>Stored the plasmid at -20 degree.</p>
+
            <h5>2016.06.07</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>2016.06.08</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>2016.06.09</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>2016.06.10</h5>
+
            <p>Extracted the plasmid, and measure the concentration.</p>
+
            <p>Stored the plasmid at -20 degree.</p>
+
 
+
 
</div>
 
</div>
 
<div class="col-md-3">
 
<div class="col-md-3">
<ul id="sidemenu">
 
<li><a href="#" onclick="toEvent('section1');">Section 1</a></li>
 
<li><a href="#" onclick="toEvent('section2');">Section 2</a></li>
 
</ul>
 
 
</div>
 
</div>
 
</div>
 
</div>

Revision as of 20:19, 9 October 2016

Notebook - iGEM NCKU

NOTEBOOK / Overview
Notebook - Overview
We divide our record - notebook into four parts:
  1. Construction
  2. Functional Test
  3. Device Design
  4. Protocols