EPF-Lausanne/22 September 2009

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==Wet Lab==
==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.
-
==People in the lab==
+
- 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
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Latest revision as of 13:45, 4 October 2009

Contents

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