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

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    <meta charset="utf-8"><link rel="shortcut icon" href="/wiki/images/8/80/T--NCKU_Tainan--favicon.png" type="image/x-icon"><link rel="icon" type="image/png" href="/wiki/images/8/80/T--NCKU_Tainan--favicon.png"><link rel="icon" type="image/x-icon" href="/wiki/images/8/80/T--NCKU_Tainan--favicon.png"><meta name="viewport" content="width=device-width, initial-scale=1.0"><meta property="og:title" content="Notebook Construction - iGEM NCKU"><meta property="og:site_name" content="Notebook Construction - iGEM NCKU"><meta property="og:description" content=""><title>Notebook Construction - iGEM NCKU</title><meta http-equiv="Content-Type" content="text/html" charset="utf-8"><meta property="og:image" content=""><meta property="og:image:type" content="image/png"><link rel="stylesheet" href="/Team:NCKU_Tainan/css/frame/T--NCKU_Tainan--bootstrap_min_css?ctype=text/css&amp;action=raw"><link href="/Team:NCKU_Tainan/font/T--NCKU_Tainan--NotoSans_css?ctype=text/css&amp;action=raw" rel="stylesheet" type="text/css"><link rel="stylesheet" href="/Team:NCKU_Tainan/font/T--NCKU_Tainan--font-awesome_min_css?ctype=text/css&amp;action=raw"> <link rel="stylesheet" href="/Team:NCKU_Tainan/css/T--NCKU_Tainan--Notebook_css?ctype=text/css&amp;action=raw">
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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">Overview</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>        </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>
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</style>
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<div class="container-fluid" style="margin-top:100px">
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<div class="head">NOTE / Construction</div>
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<div class="content row">
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<div class="col-md-9">
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<div class="head2">Notebook - Construction</div>
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<div class="title-line" id="sec1"></div>
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<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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<h5>2016.03.30</h5>
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<p>We got the bacteria with Biobrick from XMU.</p>
 +
<h5>2016.04.01</h5>
 +
<p>Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.</p>
 +
<h5>2016.04.02</h5>
 +
<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>
 +
<h5>2016.04.03</h5>
 +
<p>Extracted the plasmid, and measured the concentration.</p>
 +
<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>
 +
<p>PCR for PpI &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>
 +
<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>
 +
<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>
 +
            <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>
  
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<div class="column full_size">
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            <div class="title-line" id="sec3"></div>
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<div class="title-content">Constructed pSB1C3-B0034-K592009</div>
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<h5>2016.04.30</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>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>
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            <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>
  
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<p> Document the dates you worked on your project.</p>
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            <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>
+
            <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>
  
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<div class="column half_size">
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</div>
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<h5>What should this page have?</h5>
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<div class="col-md-3">
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<ul>
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<ul id="sidemenu">
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<li>Chronological notes of what your team is doing.</li>
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<li><a href="#" onclick="toEvent('section1');">Section 1</a></li>
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<li> Brief descriptions of daily important events.</li>
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<li><a href="#" onclick="toEvent('section2');">Section 2</a></li>
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<li>Pictures of your progress. </li>
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</ul>
−
<li>Mention who participated in what task.</li>
+
</div>
−
</ul>
+
</div>
 +
</div> <!-- /.container-fluid -->
 +
<script src="/Team:NCKU_Tainan/js/frame/T--NCKU_Tainan--jquery-1_12_0_min_js?ctype=text/javascript&amp;action=raw"></script><script src="/Team:NCKU_Tainan/js/frame/T--NCKU_Tainan--bootstrap_min_js?ctype=text/javascript&amp;action=raw"></script><script src="/Team:NCKU_Tainan/js/T--NCKU_Tainan--MathjaxConfigIgem_js?ctype=text/javascript&amp;action=raw"></script><script src="/common/MathJax-2.5-latest/MathJax.js?config=TeX-AMS-MML_HTMLorMML"></script><script src="/Team:NCKU_Tainan/js/T--NCKU_Tainan--common_js?ctype=text/javascript&amp;action=raw"></script><script>(function() {    /* change icon */    var link = document.createElement('link');    link.type = 'image/x-icon';    link.rel = 'shortcut icon';    link.href = '/wiki/images/8/80/T--NCKU_Tainan--favicon.png';    document.getElementsByTagName('head')[0].appendChild(link);}());</script>
  
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</div>
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</body></html>
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<div class="column half_size">
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<h5>Inspiration</h5>
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<p>You can see what others teams have done to organize their notes:</p>
+
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+
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<ul>
+
−
<li><a href="https://2014.igem.org/Team:ATOMS-Turkiye/Notebook">2014 ATOMS-Turkiye</a></li>
+
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<li><a href="https://2014.igem.org/Team:Tec-Monterrey/ITESM14_project.html#tab_notebook">2014 Tec Monterrey</a></li>
+
−
<li><a href="https://2014.igem.org/Team:Kyoto/Notebook/Magnetosome_Formation#title">2014 Kyoto</a></li>
+
−
<li><a href="https://2014.igem.org/Team:Cornell/notebook">2014 Cornell</a></li>
+
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</ul>
+
−
 
+
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</div>
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</html>
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Revision as of 20:19, 9 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)
2016.03.30

We got the bacteria with Biobrick from XMU.

2016.04.01

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

2016.04.02

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.

2016.04.03

Extracted the plasmid, and measured the concentration.

Store the plasmid at -20 degree.

Got the glucose sensitive promoter (PpI, Pcar) by PCR
2016.04.16

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

2016.04.17

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

Purified the PCR product and cloned to T-vector.

2016.04.18

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.

2016.04.19

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

2016.04.20

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
2016.04.30

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

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

2016.05.01

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.

2016.05.02

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

2016.05.03

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-PpI-B0034-K592009
2016.05.07

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

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

Transformed the product.

2016.05.08

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.

2016.05.09

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

2016.05.10

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)
2016.06.03

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.

2016.06.04

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.

2016.06.05

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.

2016.06.06

Extracted the plasmid, and measure the concentration.

Stored the plasmid at -20 degree.

2016.06.07

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.

2016.06.08

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.

2016.06.09

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.

2016.06.10

Extracted the plasmid, and measure the concentration.

Stored the plasmid at -20 degree.