Difference between revisions of "Team:Pasteur Paris/Protocol"

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<div id="accueilScience">
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<h1><B>Protocols</B></h1>
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<div class="text1">
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<table>
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  <thead>
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    <tr>
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      <th>Experiments</th>
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      <th>Aim:</th>
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      <th>Download</th>
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    </tr>
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  </thead>
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  <tbody>
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    <tr>
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      <td><strong><p>PCR</p></strong></td>
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      <td>To reproduce (amplify) selected sections of DNA (inserts A1, A2, B1, B2, C1, C2, E1,E2).</td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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    <tr>
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      <td><strong>Digestion</strong></td>
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      <td>To linearize the different plasmids with appropriate enzymes. Perform restriction enzyme digestion in order to recover linear backbones of the plasmids. And chose the appropriate restriction sites based on the host plasmids. </td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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    <tr>
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      <td><strong>Dephosphorylation</strong></td>
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      <td>To reduce vector self-ligation and favor that of vector to insert instead.
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</td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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    <tr>
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      <td><strong>Ligation</strong></td>
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      <td>To link an insert to its host plasmid before the transformation. This step happens after the digestion of the insert and the plasmid with the same restriction sites enzymes.
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</td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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    <tr>
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      <td><strong>Electrophoresis</strong></td>
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      <td>To know the size of DNA fragments to check the efficiency of a digestion for instance.</td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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    <tr>
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      <td><strong>Gel extraction kit</strong></td>
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      <td>To get back the DNA purified thanks to the electrophoresis on agarose gel.</td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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    <tr>
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      <td><strong>Transformation</strong></td>
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      <td>To increase the amount of plasmid by transformation in competent cells. The amount of plasmid supplied is insufficient to perform all our future experiments.</td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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    <tr>
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      <td><strong>Harvest and culture</strong></td>
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      <td>In order to obtain a large amount of plasmid, we need to grow the bacteria overnight.</td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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    <tr>
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      <td><strong>IPTG induction</strong></td>
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      <td>To increase the production of our protein which depends on this molecule.</td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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    <tr>
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      <td><strong>Purification Fast Pressure Liquid Chromatography</strong></td>
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      <td>To check if the His-tag works and if our protein has really been produced </td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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    <tr>
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      <td><strong><p>Cellulose-binding Protocol</p></strong></td>
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      <td>Verify that the C2 protein binds to cellulose.</td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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    <tr>
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      <td><strong><p>Patch protocol</p></strong></td>
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      <td> Prepare a one pot mixture that will be compressed directly after incubation.</td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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    <tr>
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      <td><strong><p>Silification & assay Protocol</p></strong></td>
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      <td> Prepare a one pot mixture that will be compressed directly after incubation.</td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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    <td><strong><p>Patch compression Protocol</p></strong></td>
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      <td>Make a patch by compression with cellulose and the silicated C2 protein.</td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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    <tr>
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      <td><strong><p>Antibody-binding Protocol</p></strong></td>
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      <td> </td>
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      <td> <a href=""><img src=""></a></td>
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    </tr>
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</table>
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        <section><center><a href="#Science"><img src="https://static.igem.org/mediawiki/2016/b/b8/SCIENCE_PASTEUR_2016.png"class="image science"  alt=""></center></a></section>
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Revision as of 21:15, 16 October 2016