Difference between revisions of "Team:Pasteur Paris/Protocol"
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<h1><B>Protocols</B></h1> | <h1><B>Protocols</B></h1> | ||
<div class="text2"> | <div class="text2"> | ||
| − | < | + | <p>During our lab work we used the following protocols. You can also download all the protocols in one file here:</p></br> |
</div> | </div> | ||
<div class="text1"> | <div class="text1"> | ||
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</thead> | </thead> | ||
<tbody> | <tbody> | ||
| + | <tr> | ||
| + | <td><strong>Microbiology equipment</strong></td> | ||
| + | <td>Materials always used in the lab </td> | ||
| + | <td> <center><a href="https://static.igem.org/mediawiki/2016/4/40/T--Pasteur_Paris--equipment.pdf"><img src="https://static.igem.org/mediawiki/2016/1/1d/T--Pasteur_Paris--DownloadT--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> | ||
| + | </tr> | ||
<tr> | <tr> | ||
<td><strong><p>PCR</p></strong></td> | <td><strong><p>PCR</p></strong></td> | ||
<td>To reproduce (amplify) selected sections of DNA (inserts A1, A2, B1, B2, C1, C2, E1,E2).</td> | <td>To reproduce (amplify) selected sections of DNA (inserts A1, A2, B1, B2, C1, C2, E1,E2).</td> | ||
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/ | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/b/be/T--Pasteur_Paris--PCR_protocol.pdf"><img src="https://static.igem.org/mediawiki/2016/1/1d/T--Pasteur_Paris--DownloadT--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><strong>Digestion</strong></td> | <td><strong>Digestion</strong></td> | ||
<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> | <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> | ||
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/ | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/a/ab/T--Pasteur_Paris--Restriction_digestion_protocol.pdf"><img src="https://static.igem.org/mediawiki/2016/1/1d/T--Pasteur_Paris--DownloadT--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><strong>Dephosphorylation</strong></td> | <td><strong>Dephosphorylation</strong></td> | ||
| − | <td>To reduce vector self-ligation and favor that of vector to insert instead. | + | <td>To reduce vector self-ligation and favor that of vector to insert instead. </td> |
| − | </td> | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/c/c0/T--Pasteur_Paris--dephosphorylation_protocol.pdf"><img src="https://static.igem.org/mediawiki/2016/1/1d/T--Pasteur_Paris--DownloadT--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> |
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/ | + | |
</tr> | </tr> | ||
<tr> | <tr> | ||
| Line 161: | Line 165: | ||
<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. | <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. | ||
</td> | </td> | ||
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/ | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/6/6f/T--Pasteur_Paris--Ligation_protocol.pdf"><img src="https://static.igem.org/mediawiki/2016/1/1d/T--Pasteur_Paris--DownloadT--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><strong>Electrophoresis</strong></td> | <td><strong>Electrophoresis</strong></td> | ||
<td>To know the size of DNA fragments to check the efficiency of a digestion for instance.</td> | <td>To know the size of DNA fragments to check the efficiency of a digestion for instance.</td> | ||
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/ | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/6/60/T--Pasteur_Paris--Gel_electrophoresis_protocol.pdf"><img src="https://static.igem.org/mediawiki/2016/1/1d/T--Pasteur_Paris--DownloadT--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><strong>Gel extraction kit</strong></td> | <td><strong>Gel extraction kit</strong></td> | ||
<td>To get back the DNA purified thanks to the electrophoresis on agarose gel.</td> | <td>To get back the DNA purified thanks to the electrophoresis on agarose gel.</td> | ||
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/ | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/4/4e/T--Pasteur_Paris--Gel_extraction_protocol.pdf"><img src="https://static.igem.org/mediawiki/2016/1/1d/T--Pasteur_Paris--DownloadT--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><strong>Transformation</strong></td> | <td><strong>Transformation</strong></td> | ||
<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> | <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> | ||
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/ | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/c/ca/T--Pasteur_Paris--Transformation_protocol.pdf"><img src="https://static.igem.org/mediawiki/2016/1/1d/T--Pasteur_Paris--DownloadT--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> |
| + | </tr> | ||
| + | <tr> | ||
| + | <td><strong>Miniprep QIAGEN Kit</strong></td> | ||
| + | <td> To extract DNA from a culture of bacteria</td> | ||
| + | <td> <center><a href="https://static.igem.org/mediawiki/2016/d/d5/T--Pasteur_Paris--Miniprep_protocol.pdf"><img src="https://static.igem.org/mediawiki/2016/1/1d/T--Pasteur_Paris--DownloadT--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><strong>Harvest and culture</strong></td> | <td><strong>Harvest and culture</strong></td> | ||
<td>In order to obtain a large amount of plasmid, we need to grow the bacteria overnight.</td> | <td>In order to obtain a large amount of plasmid, we need to grow the bacteria overnight.</td> | ||
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/ | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/4/4b/T--Pasteur_Paris--Bacterial_culture_protocol.pdf"><img src="https://static.igem.org/mediawiki/2016/1/1d/T--Pasteur_Paris--DownloadT--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> |
| + | </tr> | ||
| + | <tr> | ||
| + | <td><strong>Stab culture</strong></td> | ||
| + | <td>To store a clone of bacteria to be used later.</td> | ||
| + | <td> <center><a href="https://static.igem.org/mediawiki/2016/b/b1/Stab_culture_Pasteur_Paris_2016.pdf"><img src="https://static.igem.org/mediawiki/2016/1/1d/T--Pasteur_Paris--DownloadT--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><strong>IPTG induction</strong></td> | <td><strong>IPTG induction</strong></td> | ||
<td>To increase the production of our protein which depends on this molecule.</td> | <td>To increase the production of our protein which depends on this molecule.</td> | ||
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/ | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/a/a4/T--Pasteur_Paris--Protein_induction_protocol.pdf"><img src="https://static.igem.org/mediawiki/2016/1/1d/T--Pasteur_Paris--DownloadT--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td><strong>Purification Fast | + | <td><strong>Protein Purification Fast Protein Liquid Chromatography</strong></td> |
<td>To check if the His-tag works and if our protein has really been produced </td> | <td>To check if the His-tag works and if our protein has really been produced </td> | ||
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/ | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/0/07/T--Pasteur_Paris--FPLC_Protein_purification_protocol.pdf"><img src="https://static.igem.org/mediawiki/2016/1/1d/T--Pasteur_Paris--DownloadT--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><strong><p>Cellulose-binding Protocol</p></strong></td> | <td><strong><p>Cellulose-binding Protocol</p></strong></td> | ||
<td>Verify that the C2 protein binds to cellulose.</td> | <td>Verify that the C2 protein binds to cellulose.</td> | ||
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/1/ | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/e/e1/Protocol-cellulose-binding_Pasteur_Paris2016.pdf"><img src="https://static.igem.org/mediawiki/2016/1/15/T--Pasteur_Paris--Chemistry_T--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> |
</tr> | </tr> | ||
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<td><strong><p>Silification & assay Protocol</p></strong></td> | <td><strong><p>Silification & assay Protocol</p></strong></td> | ||
<td> Prepare a one pot mixture that will be compressed directly after incubation.</td> | <td> Prepare a one pot mixture that will be compressed directly after incubation.</td> | ||
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/1/ | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/2/2a/Protocol-silification-assay_Pasteur_Paris2016.pdf"><img src="https://static.igem.org/mediawiki/2016/1/15/T--Pasteur_Paris--Chemistry_T--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> |
| − | + | ||
</tr> | </tr> | ||
| + | <tr> | ||
| + | <td><strong><p>Model of the mechanical properties of the patch</p></strong></td> | ||
| + | <td>Predict the influence of silification on the mechanical properties of the patch</td> | ||
| + | <td> <center><a href="https://static.igem.org/mediawiki/2016/d/da/T--Pasteur_Paris--Mechanical_patch_properties_protocol.pdf"><img src="https://static.igem.org/mediawiki/2016/7/76/T--Pasteur_Paris--Patch_T--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> | ||
| + | </tr> | ||
| + | <tr> | ||
<td><strong><p>Patch compression Protocol</p></strong></td> | <td><strong><p>Patch compression Protocol</p></strong></td> | ||
<td>Make a patch by compression with cellulose and the silicated C2 protein.</td> | <td>Make a patch by compression with cellulose and the silicated C2 protein.</td> | ||
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/ | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/a/ac/Protocol-patch-compression_Pasteur_Paris.pdf"><img src="https://static.igem.org/mediawiki/2016/7/76/T--Pasteur_Paris--Patch_T--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td><strong><p>Patch protocol</p></strong></td> | + | <td><strong><p>One pot mixture-Patch protocol</p></strong></td> |
<td> Prepare a one pot mixture that will be compressed directly after incubation.</td> | <td> Prepare a one pot mixture that will be compressed directly after incubation.</td> | ||
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/ | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/8/89/Protocol-one-pot-mixture_Pasteur_Paris2016.pdf"><img src="https://static.igem.org/mediawiki/2016/7/76/T--Pasteur_Paris--Patch_T--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><strong><p>Antibody-binding Protocol</p></strong></td> | <td><strong><p>Antibody-binding Protocol</p></strong></td> | ||
| − | <td> </td> | + | <td> The goal is to determine whether our immunoassay is effective. We want to ensure that our antibodies are well able to detect the presence or absence of viral proteins from Chikungunya and Yellow Fever viruses. |
| − | <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/ | + | </td> |
| − | + | <td> <center><a href="https://static.igem.org/mediawiki/2016/2/21/T--Pasteur_Paris--Immunoassay_Protocol.pdf"><img src="https://static.igem.org/mediawiki/2016/f/f1/T--Pasteur_Paris--Antibody_Pasteur_PARIS_2016.png" width="40%" alt=""></a></center></td> | |
| − | + | ||
</tr> | </tr> | ||
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</br></br></br> | </br></br></br> | ||
</div> | </div> | ||
| − | + | <div class="text2"> | |
| + | <p></a> | ||
| + | You can also download all the protocols in one file here:</br></br> | ||
| + | <center><a href="https://static.igem.org/mediawiki/2016/6/66/T--Pasteur_Paris--Protocols.pdf"><img src="https://static.igem.org/mediawiki/2016/3/3a/T--Pasteur_Paris--Protocols_Pasteur_Paris_2016.png" width="10%" alt=""></a></center></br></br></br></br></br></br></p> | ||
| + | </div> | ||
<div class="text1"> | <div class="text1"> | ||
| − | <section><center><a href="#Science"><img src="https://static.igem.org/mediawiki/2016/ | + | <section><center><a href="#Science"><img src="https://static.igem.org/mediawiki/2016/c/c7/T--Pasteur_Paris--SCIENCE_PASTEUR_2016.png"class="image science" alt=""></center></a></section> |
</div> | </div> | ||
