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<h1><B>Protocols</B></h1> | <h1><B>Protocols</B></h1> | ||
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| − | During our lab work we used the following protocols. You can also download them here:<a href=""><img src=""></a>. | + | </a>During our lab work we used the following protocols. You can also download them here:<a href=""><img src=""></a>. |
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| 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 |
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