EPF-Lausanne/6 October 2009

Wet Lab

2nd gel

We did a second gel of yesterday's colony PCR to check the gel as iti is strange not to have any double transformant but we had double antibiotic selection.

New colony PCR

We think that maybe if the first colony PCR didn't show any double transformants it is because we didn't let enough time for the extansion (1'20) and RO2 is 1900 bp long and Taq only goes at 1 kb / minute. From now, we will let an extansion time up to 2 minutes.

Replating

... of RO2.4 + BB1 (3 Trp K.O. strains), RO1.1 + BB1 (3 strains).

Gel

Still no double transformants !!!

Culture

Prepared cultures of 2 x 7mL of DH5 alpha RO2.4 + BB1 clone n.3 (the strain that works) in LB for tomorrow's experiment (the cells were taken from the glycerol stock). Left cells to incubate overnight at 37°C.

People in the lab

Gab, Christian