EPF-Lausanne/15 October 2009

Wet Lab

Did an experiment to test the minimal induction time of our LovTap:

• We grew an overnight culture of DH5-alpha RO2.4+BB1 (yesterday)

• Took Erlens of 25 mL and inoculated 0.5 mL of culture into 3.5 mL of fresh LB, with 2mM IPTG.

• Kept tubes in the dark, and put one Erlen into the incubator with blue light every 5 min.

• Did that for 5, 10, 15, ..., 50 min.

• After 50 min, took all Erlens out, loaded 200 ul of the culture into each well (loaded 4 different wells per Erlen) and took the measurements of fluorescence and OD with the plate reader.

The resulting graph is:

Results

Also, we prepared the DNA of all our parts to send it to the Registry. DNA was sent out in the end of the afternoon and should arrive on Monday at the latest.

Used mutagenesis kit to mutate either ILE427PHE or LEU453GLY on the LovTap protein: this is to test whether the mutated LovTap is more stable in its light state than the original LovTap. Cells were transformed with the new mutated plasmids.

People in the lab

Heidi, Gab, Tu, Basile