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

Revision as of 21:42, 16 October 2016

Protocols

During our lab work we used the following protocols. You can also download them here:

Experiments	Aim:	Download
PCR	To reproduce (amplify) selected sections of DNA (inserts A1, A2, B1, B2, C1, C2, E1,E2).
Digestion	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.
Dephosphorylation	To reduce vector self-ligation and favor that of vector to insert instead.
Ligation	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.
Electrophoresis	To know the size of DNA fragments to check the efficiency of a digestion for instance.
Gel extraction kit	To get back the DNA purified thanks to the electrophoresis on agarose gel.
Transformation	To increase the amount of plasmid by transformation in competent cells. The amount of plasmid supplied is insufficient to perform all our future experiments.
Harvest and culture	In order to obtain a large amount of plasmid, we need to grow the bacteria overnight.
IPTG induction	To increase the production of our protein which depends on this molecule.
Purification Fast Pressure Liquid Chromatography	To check if the His-tag works and if our protein has really been produced
Cellulose-binding Protocol	Verify that the C2 protein binds to cellulose.
Patch protocol	Prepare a one pot mixture that will be compressed directly after incubation.
Silification & assay Protocol	Prepare a one pot mixture that will be compressed directly after incubation.
Patch compression Protocol	Make a patch by compression with cellulose and the silicated C2 protein.
Antibody-binding Protocol

@@ Line 207: / Line 207: @@
        <td><strong><p>Silification & assay Protocol</p></strong></td>
        <td> Prepare a one pot mixture that will be compressed directly after incubation.</td>
-       <td> <a href=""><img src=""></a></td>
+       <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/2/26/IGEM16_Pasteur_Paris_silica.png" width="40%" alt=""></a></center></td>
      </tr>
       <td><strong><p>Patch compression Protocol</p></strong></td>
        <td>Make a patch by compression with cellulose and the silicated C2 protein.</td>
-       <td> <a href=""><img src=""></a></td>
+       <td> <center><a href=""><img src="https://static.igem.org/mediawiki/2016/2/2d/Patch_T--iGEM16_Pasteur_Paris--file.xxx.png" width="40%" alt=""></a></center></td>
      </tr>