Difference between revisions of "Team:EPF Lausanne/Notebook/Yeast"
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<li>Restriction analysis</li> | <li>Restriction analysis</li> | ||
<li>Polymerase Chain Reaction</li> | <li>Polymerase Chain Reaction</li> | ||
| − | <li> | + | <li>Yeast integration</li> |
</ul> | </ul> | ||
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| − | <p><small>We used four different sets of enzymes for the restriction analysis. Linearized pTPGI_dCas9_VP64 is expected to be 10'987 bp. We observe that the gel corresponds to the expected one (fig. 2). </br> | + | <p><small>We used four different sets of enzymes for the restriction analysis. Linearized pTPGI_dCas9_VP64 is expected to be 10'987 bp. We observe that the gel (fig.1) corresponds to the expected one (fig. 2). </br> |
The plasmid was linearised both with EagI HF and NotI HF prior to integration. We integrated each linearised plasmid to obtain two different yeast strains. | The plasmid was linearised both with EagI HF and NotI HF prior to integration. We integrated each linearised plasmid to obtain two different yeast strains. | ||
</small></p> | </small></p> | ||
Revision as of 17:12, 17 August 2015
saccharomyces cerevisiae
Integrate pTPGI_dCas9_VP64Integrate pTPGI_dCas9_VP64
We received plasmid pTPGI_dCas9_VP64 in bacteria from Addgene. We inoculated single colonies of bacteria in order to prepare glycerol stocks and minipreps. We performed a restriction analysis to check the identity of our plasmid. We linearised the plasmid by PCR, in order to integrate it into yeast genome.
Materials and method
- Glycerol stocks
- Miniprep
- Restriction analysis
- Polymerase Chain Reaction
- Yeast integration
Results
We used four different sets of enzymes for the restriction analysis. Linearized pTPGI_dCas9_VP64 is expected to be 10'987 bp. We observe that the gel (fig.1) corresponds to the expected one (fig. 2). The plasmid was linearised both with EagI HF and NotI HF prior to integration. We integrated each linearised plasmid to obtain two different yeast strains.
