EPF-Lausanne/8 September 2009

Wet Lab

Cultures

Results:

RO1 has grown in LB and M9

RO2 has grown in LB but not in M9

RO2+BB #1,4 did not grow

RO2+BB #3 has grown in LB but not in M9

-> problem with our glycerol stock (RO2 + BB didn't grow in LB !!). We did some plates of the cultures that grew.

For safety we did some plates with the glycerol stocks of RO2 + BB 1,4,8. We also did some plates with the cultures that grew (RO1 1,2,3, RO2 4,5,10, RO2 + BB 3).

Characterization

As RO1 grew in both media, we put it in 25mL medium with Amp. We did also with RO2 (LB), and RO2+BB (LB), and LacI-RFP.

For the characterisation, we did different combinations : -TRP, -TRP -ATC, +TRP -ATC, +TRP +ATC. Evertime for all the clones in M9 and then in LB.

TRP : LB : dilution 50x so 1ul, M9 : dilution 25x so 2 ul. We took the stock solution 4,250 g/L.

ATC : 5x dilution so 10 ul from the 500 ng/ml stock solution.

to be completed ...

People in the lab

Mélanie, Caroline, Basile, Nicolas