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

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<h1>Notebook</h1>
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      <div id="top">
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<p> Document the dates you worked on your project. This should be a detailed account of the work done each day for your project.</p>
+
      </div>
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      <div id="category" class="vertical-container">
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          <h1 class="wet">Notebook</h1>
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      </div>
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    </div>
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  </div>
 
</div>
 
</div>
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<div class="clear"></div>
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<div class="container-fluid">
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  <div class="row">
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    <div id="paragraph" class="paragraph col-md-8 col-md-offset-1 col-xs-offset-1 col-xs-10">
 +
      <h2 id="BBa_K2275007">Construction of BBa_K2275007</h2>
 +
      <hr>
 +
      <h3>5/22</h3>
 +
      <ul>
 +
        <li>
 +
          PCR to get DNA of <i>nir</i> gene cluster from <i>Eschericha coli</i> MG1655 and analyze by DNA gel.
 +
        </li>
 +
      </ul>
 +
      <h3>5/23</h3>
 +
      <ul>
 +
        <li>
 +
          Digest <i>nir</i> gene cluster and pSB1C3-P<sub>LacI</sub>-sfGFP with <i>Hind</i>Ⅲ and <i>SpeI</i>. Analyze by the DNA gel.
 +
        </li>
 +
        <li>
 +
          Ligase at 16℃ overnight.
 +
        </li>
 +
      </ul>
 +
      <h3>5/25</h3>
 +
      <ul>
 +
        <li>
 +
          Transform the ligased product into DH5&alpha; and put at 37℃ for 12 hours.
 +
        </li>
 +
      </ul>
 +
      <h3>5/26</h3>
 +
      <ul>
 +
        <li>
 +
          Select five colonies from the plates and pre-culturing.
 +
        </li>
 +
      </ul>
 +
      <h3>5/27</h3>
 +
      <ul>
 +
        <li>
 +
          Extract plasmid and use plasmid digestion to confirm.
 +
        </li>
 +
      </ul>
  
 +
      <h2 id="BBa_K2275008">Construction of BBa_K2275008</h2>
 +
      <hr>
 +
      <h3>7/3</h3>
 +
      <ul>
 +
        <li>
 +
        PCR to get DNA of <i>gudB</i> from <i>Bacillus subtilis</i> and analyze by the DNA gel.
 +
        </li>
 +
        <li>
 +
          Digest gene <i>gudB</i> and pSB1C3-P<sub>LacI</sub>-sfGFP with BamHI and PstI. Analyze by the DNA gel
 +
        </li>
 +
        <li>
 +
          Ligase at 16℃ overnight.
 +
        </li>
 +
      </ul>
 +
      <h3>7/4</h3>
 +
      <ul>
 +
        <li>
 +
          Transform the ligased product into DH5&alpha; and put at 37℃ for 12 hours.
 +
        </li>
 +
      </ul>
 +
      <h3>7/5</h3>
 +
      <ul>
 +
        <li>
 +
          Confirmation by colony PCR.
 +
        </li>
 +
        <li>
 +
          Select two colonies from the plates and pre-culturing.
 +
        </li>
 +
      </ul>
 +
      <h3>7/6</h3>
 +
      <ul>
 +
        <li>
 +
          Extract plasmid and use plasmid digestion to confirm.
 +
        </li>
 +
      </ul>
  
−
<div class="column half_size">
+
      <h2 id="BBa_K2275009">Construction of BBa_K2275009</h2>
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<h5>What should this page have?</h5>
+
      <hr>
−
<ul>
+
      <<h3>8/13</h3>
−
<li>Chronological notes of what your team is doing.</li>
+
      <ul>
−
<li> Brief descriptions of daily important events.</li>
+
        <li>
−
<li>Pictures of your progress. </li>
+
          PCR to get DNA of <i>gudB</i> from <i>Pseudomonos putida</i> and analyze by the DNA gel.
−
<li>Mention who participated in what task.</li>
+
        </li>
−
</ul>
+
        <li>
−
 
+
          Digest gene glnA and pSB1C3-P<sub>LacI</sub>-sfGFP with <i>Hind</i>Ⅲ and PstI. Analyze by the DNA gel
 +
        </li>
 +
        <li>
 +
          Ligase at 16℃, 4 hours and then transform into DH5&alpha;.
 +
        </li>
 +
      </ul>
 +
      <h3>8/14</h3>
 +
      <ul>
 +
        <li>
 +
          Select two colonies from the plates and pre-culturing.
 +
        </li>
 +
        <li>
 +
          Extract plasmid and use plasmid digestion to confirm.
 +
        </li>
 +
      </ul>
 +
      <h2 id="BBa_K2275010">Construction of BBa_K2275010</h2>
 +
      <hr>
 +
      <h3>9/6</h3>
 +
      <ul>
 +
        <li>PCR to get rev-<i>gudB</i>.</li>
 +
        <li>Digest gene rev-<i>gudB</i> with XbaI and PstI and pSB1C3-P<sub>LacI</sub>-glnA with <i>Spe</i>I and <i>Pst</i>I. Analyze by the DNA gel.</li>
 +
        <li>Ligase at 16℃ , 4 hours and then transform into DH5&alpha;</li>
 +
      </ul>
 +
      <h3>9/7</h3>
 +
      <ul>
 +
        <li>
 +
          Select two colonies from the plates and pre-culturing.
 +
        </li>
 +
      </ul>
 +
      <h3>9/8</h3>
 +
      <ul>
 +
        <li>
 +
          Extract plasmid and confirm with plasmid digestion.
 +
        </li>
 +
      </ul>
 +
      <h2 id="BBa_K2275011">Construction of BBa_K2275011</h2>
 +
      <hr>
 +
      <h3>9/13</h3>
 +
      <ul>
 +
        <li>
 +
          Add a NsrR binding site to K381001 before RBS by PCR and analyze by DNA gel.
 +
        </li>
 +
      </ul>
 +
      <h3>9/14</h3>
 +
      <ul>
 +
        <li>Digest linear PCR product with <i>Dpn</i>I and self-ligate by phosphorylation</li>
 +
        <li>Transform the ligased product into DH5&alpha; and put at 37℃ for 12 hours.</li>
 +
      </ul>
 +
      <h3>9/15</h3>
 +
      <ul>
 +
        <li>
 +
          Select two colonies from the plates and pre-culturing.
 +
        </li>
 +
      </ul>
 +
      <h3>9/16</h3>
 +
      <ul>
 +
        <li>
 +
          Extract plasmid and use plasmid digestion to confirm.
 +
        </li>
 +
      </ul>
 +
      <h2 id="BBa_K2275012">Construction of BBa_K2275012</h2>
 +
      <hr>
 +
      <h3>9/13</h3>
 +
      <ul>
 +
        <li>
 +
          Add a NsrR binding site to K381001 after RBS by PCR and analyze by DNA gel.
 +
        </li>
 +
      </ul>
 +
      <h3>9/14</h3>
 +
      <ul>
 +
        <li>
 +
        Digest the linear PCR product with <i>Dpn</i>I and self-ligate by phosphorylation.
 +
        </li>
 +
        <li>
 +
          Transform the ligased product into DH5&alpha;  and put at 37℃ for 12 hours
 +
        </li>
 +
      </ul>
 +
      <h3>9/15</h3>
 +
      <ul>
 +
        <li>
 +
          Select two colonies from the plates and pre-culturing.
 +
        </li>
 +
      </ul>
 +
      <h3>9/16</h3>
 +
      <ul>
 +
        <li>
 +
          Extract plasmid and use plasmid digestion to confirm.
 +
        </li>
 +
      </ul>
 +
      <h2 id="sensing_test">Nitrate sensing test</h2>
 +
      <hr>
 +
      <h3>7/1~7/31</h3>
 +
      <ul>
 +
        <li>
 +
          Use M2 to quantitate fluorescence emitting by K381001 in lyophilized <i>E. coli</i> powder
 +
        </li>
 +
      </ul>
 +
      <h3>8/1~8/31</h3>
 +
      <ul>
 +
        <li>
 +
          Use our device to do the nitrate sensing test by K381001 in lyophilized <i>E. coli</i> powder.
 +
        </li>
 +
      </ul>
 +
      <h2 id="pathway_test">Whole pathway test</h2>
 +
      <hr>
 +
      <h3>10/7~10/10</h3>
 +
      <ul>
 +
        <li>
 +
          Use our device and modified <i>E. coli</i> to test our whole project.
 +
        </li>
 +
      </ul>
 +
    </div>
 +
    <div id="sidemenu" class="col-md-2">
 +
      <div class="list-group">
 +
        <a onclick="scrollto('#BBa_K2275007')" class="list-group-item">BBa_K2275007</a>
 +
        <hr>
 +
        <a onclick="scrollto('#BBa_K2275008')" class="list-group-item">BBa_K2275008</a>
 +
        <hr>
 +
        <a onclick="scrollto('#BBa_K2275009')" class="list-group-item">BBa_K2275009</a>
 +
        <hr>
 +
        <a onclick="scrollto('#BBa_K2275010')" class="list-group-item">BBa_K2275010</a>
 +
        <hr>
 +
        <a onclick="scrollto('#BBa_K2275011')" class="list-group-item">BBa_K2275011</a>
 +
        <hr>
 +
        <a onclick="scrollto('#BBa_K2275012')" class="list-group-item">BBa_K2275012</a>
 +
        <hr>
 +
        <a onclick="scrollto('#sensing_test')" class="list-group-item">Nitrate sensing test</a>
 +
        <hr>
 +
        <a onclick="scrollto('#pathway_test')" class="list-group-item">Whole pathway test</a>
 +
        <hr>
 +
        <a class="list-group-item top"><i  onclick="scrollto('#top')" class="fa fa-arrow-up fa-1x" aria-hidden="true"></i></a>
 +
      </div>
 +
    </div>
 +
  </div>
 
</div>
 
</div>
  
−
<div class="column half_size">
 
−
<h5>Inspiration</h5>
 
−
<p>You can see what others teams have done to organize their notes:</p>
 
  
−
<ul>
 
−
<li><a href="https://2014.igem.org/Team:ATOMS-Turkiye/Notebook">2014 ATOMS-Turkiye</a></li>
 
−
<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>
 
−
</ul>
 
  
−
</div>
 
 
</html>
 
</html>

Latest revision as of 20:41, 31 October 2017

Notebook

Construction of BBa_K2275007

5/22

  • PCR to get DNA of nir gene cluster from Eschericha coli MG1655 and analyze by DNA gel.

5/23

  • Digest nir gene cluster and pSB1C3-PLacI-sfGFP with HindⅢ and SpeI. Analyze by the DNA gel.
  • Ligase at 16℃ overnight.

5/25

  • Transform the ligased product into DH5α and put at 37℃ for 12 hours.

5/26

  • Select five colonies from the plates and pre-culturing.

5/27

  • Extract plasmid and use plasmid digestion to confirm.

Construction of BBa_K2275008

7/3

  • PCR to get DNA of gudB from Bacillus subtilis and analyze by the DNA gel.
  • Digest gene gudB and pSB1C3-PLacI-sfGFP with BamHI and PstI. Analyze by the DNA gel
  • Ligase at 16℃ overnight.

7/4

  • Transform the ligased product into DH5α and put at 37℃ for 12 hours.

7/5

  • Confirmation by colony PCR.
  • Select two colonies from the plates and pre-culturing.

7/6

  • Extract plasmid and use plasmid digestion to confirm.

Construction of BBa_K2275009

<

8/13

  • PCR to get DNA of gudB from Pseudomonos putida and analyze by the DNA gel.
  • Digest gene glnA and pSB1C3-PLacI-sfGFP with HindⅢ and PstI. Analyze by the DNA gel
  • Ligase at 16℃, 4 hours and then transform into DH5α.

8/14

  • Select two colonies from the plates and pre-culturing.
  • Extract plasmid and use plasmid digestion to confirm.

Construction of BBa_K2275010

9/6

  • PCR to get rev-gudB.
  • Digest gene rev-gudB with XbaI and PstI and pSB1C3-PLacI-glnA with SpeI and PstI. Analyze by the DNA gel.
  • Ligase at 16℃ , 4 hours and then transform into DH5α

9/7

  • Select two colonies from the plates and pre-culturing.

9/8

  • Extract plasmid and confirm with plasmid digestion.

Construction of BBa_K2275011

9/13

  • Add a NsrR binding site to K381001 before RBS by PCR and analyze by DNA gel.

9/14

  • Digest linear PCR product with DpnI and self-ligate by phosphorylation
  • Transform the ligased product into DH5α and put at 37℃ for 12 hours.

9/15

  • Select two colonies from the plates and pre-culturing.

9/16

  • Extract plasmid and use plasmid digestion to confirm.

Construction of BBa_K2275012

9/13

  • Add a NsrR binding site to K381001 after RBS by PCR and analyze by DNA gel.

9/14

  • Digest the linear PCR product with DpnI and self-ligate by phosphorylation.
  • Transform the ligased product into DH5α and put at 37℃ for 12 hours

9/15

  • Select two colonies from the plates and pre-culturing.

9/16

  • Extract plasmid and use plasmid digestion to confirm.

Nitrate sensing test

7/1~7/31

  • Use M2 to quantitate fluorescence emitting by K381001 in lyophilized E. coli powder

8/1~8/31

  • Use our device to do the nitrate sensing test by K381001 in lyophilized E. coli powder.

Whole pathway test

10/7~10/10

  • Use our device and modified E. coli to test our whole project.