EPF-Lausanne/21 August 2009

Wet Lab

Miniprep of the 2 possible readout 1 plasmids.

Colony PCR of the LacI-RBS-LovTap-Term that grew to check whether the insert is correct. Then agarose gel to verifiy length of fragment.

Also, since we wanted to do further tests on the maybe readout 1 plasmids, we did a classic PCR with the iGEM primers on the plasmids, and a digestion assay with SpeI (since the TrpOperon has 2 SpeI sites in its sequence). Checked the results on the gel as well.

Results of the first characterization

Experiment on Read Out 2. Only the clones 4, 5 and 10 had nice curves. The experiment was made using LB. For every clone there was three different possibilities : without anything OR with tryptophan OR with ATC. The cells had all exactly the same initial conditions, the products were added only at the last moment. We can see that clone 4 and 10 show a very good answer to ATC as well as to TRP.

Clone 4 :

Clone 5 :

Clone 10 :

People in the lab

Basile, Gab, Christian