Difference between revisions of "Team:EPF Lausanne/Notebook/Yeast"
| Line 40: | Line 40: | ||
<ul> | <ul> | ||
<li <a href="#Integrate_reporter_plasmid">Integrate reporter plasmid</a></li> | <li <a href="#Integrate_reporter_plasmid">Integrate reporter plasmid</a></li> | ||
| + | </u1> | ||
<li><a href="#synthesize_promoters">Synthesize promoters</a></li> | <li><a href="#synthesize_promoters">Synthesize promoters</a></li> | ||
<li><a href="#PCRoverlaps_prom">Add Gibson overlaps to CYC promoters by PCR</a></li> | <li><a href="#PCRoverlaps_prom">Add Gibson overlaps to CYC promoters by PCR</a></li> | ||
<li><a href="#gibsonprom">Gibson assembly of promoters in reporter plasmid</a></li> | <li><a href="#gibsonprom">Gibson assembly of promoters in reporter plasmid</a></li> | ||
<li><a href="#integrate_reporters_plasmids">Integrate the reporter plasmids</a></li> | <li><a href="#integrate_reporters_plasmids">Integrate the reporter plasmids</a></li> | ||
| − | |||
| − | |||
<ul> | <ul> | ||
| + | <li><a href="#Integrate_and_express_gRNAs">Integrate and express gRNAs</a></li> | ||
| + | </ul> | ||
<li><a href="#synthesize_gRNAs">Synthesize gRNAs</a></li> | <li><a href="#synthesize_gRNAs">Synthesize gRNAs</a></li> | ||
<li><a href="#PCRoverlaps_gRNAs">Add Gibson overlaps to gRNAs by PCR</a></li> | <li><a href="#PCRoverlaps_gRNAs">Add Gibson overlaps to gRNAs by PCR</a></li> | ||
Revision as of 14:20, 17 August 2015
saccharomyces cerevisiae
pTPGI_dCas9_VP64 integration
As we need dCas9 expressing yeasts, we decided to integrate a plasmid that would inducively express dCas9.
First and second trial
The negative and positive control worked fine, but the transformation control didn’t work. After investigation we found out that we didn’t have an origin of replication in our plasmid thus not enabling our yeasts to replicate the plasmid and to grow. See more details here
