EPF-Lausanne/25 September 2009

Wet Lab

Colony PCR

- 5 clones RO2#4 + BB1 JRG 1046

- 4 clones RO1#1 + BB1 JW 4356-2

- 1 clones RO1#1 JRG 465

- 5 clones RO1#1 + BB1 JRG 1046

- Negative control

Numerotation of the samples :

1 : RO1#1 + BB1 JW 4356-2

2 : RO1#1 JRG 456

3 : RO1#1 + BB1 JRG 1046

4 : RO2#4 + BB1 JRG 1046

Counting of colonies for tests of competent cells

- JW 4356-2 : bacterial lawn -> the cells are well competent.

- JRG 1046 : 100pg : 49x4 -> 196 cells. Good efficacy : 10^6 - 10^8

- JRG 465 : 10 pg : about 100 cells. 1ug -> 10^5 -> 10^7 so good efficacy as well.

Gel

We loaded samples 1,2,3,4, to see if the bands are at the right places. Conclusion : RO1#1 + BB1 are okay and RO1#1 + BB1 as well but RO1#1 and RO2#4 + BB1 didn't work.

Inoculation of cells for chance

-RO1-BB1 #2 JRG1046 +IPTG/-IPTG

-RO1-BB1 #1 JW4356-2 +IPTG/-IPTG

-RO1#1-BB1 #3 JW 4356-2 +IPTG/-IPTG

-RO1#1-BB1 #5 JRG 1046 +IPTG/-IPTG

-RO1#1 JW4356-2

-RO2#4 JRG 465

-RO2#4 JW4356-2

All of them 700 ul in 5 ml of LB.

People in the lab

Gab, Nicolas