EPF-Lausanne/20 August 2009

Wet Lab

Long day.......

Miniprep of the LovTap-Term clones we'd put in liquid culture. Then we tried to do the whole protocol to obtain our BioBrick LacI-RBS-LovTap-Term by 2 strategies:

- classic iGEM protocol: amplify LacI-RBS by PCR, purify, then digest it with E/S and digest the LovTap-Term vector with E/X , dephosphorylate the vector, purify again, then do the ligations of the various LacI-RBS with the LovTap-Terms in random combinations

- the "dirty" technique: digest both the LovTap-Term and LacI-RBS plasmids, dephosphorylate the LovTap-Term vector, purify, then do the ligation by just "mixing" the 2 solutions (so containing pieces of both vectors). The point is that since the LovTap-Term vector has a kanamycin resistance and the LacI-RBS is ampicilin, only the correct combination should manage to grow

We then did the transformations with the resulting ligation products. Plates were left to incubate overnight at 37°C.

Also, we obtained 2 clones from the previous day transformations with readout system 1. Did a colony PCR and a gel to check the insert, but the result of the gel showed some bands which we couldn't seem to identify. Did liquid cultures of the 2 clones so that we can do more tests tomorrow.

People in the lab

Basile, Gab, Christian, Heidi