EPF-Lausanne/17 October 2009

Wet Lab

Miniprep of clones n°1 and n°10 containing the plasmid with the gene for the mutated LovTap.

Transformed Dh5-alpha competent cells with:

• RO1.1
• RO2.4
• RO1.1 + BB1
• RO2.4 + BB1
• RO1.1 + mutated BB10
• RO2.4 + mutated BB10

This is to have "new" cells with our constructs, since somehow double-transformants sometimes appear to lose one of the plasmids or something else, or they don't work as effectively as they did earlier. Maybe this'll solve our problems?

Redid the experiment we did on Thursday, so putting one Erlen of cells with RO2.4 + BB1 into the experimental conditions every 5 min. This time we'll keep one tube in the dark, so that we see the "basal" level of RFP expression:

RO2 double-transformants in DH5-alpha

Took the picture of the polyacrylamide gel that Basile did yesterday, to see whether the LacI promoter we'd taken from the kit and used for our LovTap biobrick works: we wanted to see whether there's a different level of expression of our LovTap between cells with or without IPTG added to the medium. In theory, the +IPTG should have a higher level of expression than the -IPTG. From the gel we can see that the patterns of expression as exactly the same --> IPTG has no influence on the level of expression of LovTap.

People in the lab

Gab, Tu, Christian