EPF-Lausanne/22 September 2009

Wet Lab

Results of the cultures after one more night

- RO1 + BB double transformants grew (white).

- RO2 #4,5,10 didn't grow.

- RO1 #1,2 didn't grow.

- RO2 + BB double transformatnts grew. #8 is pink, #3 is white and # 7 is pink.

Trp K.O. strains

After about 35-40 hours.

2 out of 3 grew : JW 4356-2 0.14, JRG 1046 0.00, JRG 465 : 0.04...

We will let them more time at 18°C... maybe it will grow a bit more !!

New cultures

TrpR K.O.

-JW4356-2 (KANA)

-JRG1046

- JRG 465

All the 3 in 3ml of SOB, at 37°C. We took them from the glycerol stock.

Cultures for characterization

IPTG : stock solution of 100 mM. Induction at 1 mM should be enough, so we have to dilute it 100x.

- RO2 #4 - BB1, clone #3. 4 conditions : +IPTG +light, +IPTG -light, -IPTG -light, -IPTG +light. Antibiotic : 10 ul amp + 30 ul kana.

- RO1#1 -BB1, clone #1. The same 4 conditions as above. Antibio : 10 ul amp + 30 ul kana.

- Negative control : BB1 (BB1.13). Antibio : 30 ul Kana.

All in 5 ml M9/minimal + AA + thiamine + corresponding antibiotic.

People in the lab

Basile