EPF-Lausanne/24 July 2009

Wet Lab

We found a Term plasmid that last year team used, and that worked : BBa-B0015.

We did 3 ligations in parallel :

LovTAP-Term with iGEM protocol and using the plasmid BBa-B0015.

LovTAP-Term using the Term digested last year with EX. (The idea is to insert LovTAP in the Term directly)

LacI-RBS with iGEM protocol again.

So we first did the digestion :

1. LovTAP cut with E and S, Term plasmid cut with X and P, the backbone cut with E and P.

2. LovTAP cut with E and S. We will use the Term from last year, already digested.

3. LacI with E and S, RBS with X and P and the backbone with E and P.

Next we ran a gel to check the digestion products.

We did the 3 ligations in parallel.

At this time we had the results of the gel, and actually the digestion didn't work. The backbones were digested but not the rest. There were no bands at all for the LovTAP and the others weren't digested. We will begin all over but using another protocol than iGEM's for the digestion.

People in the lab

Nathalie, Caroline, Mélanie, Tu, Heidi