EPF-Lausanne/22 September 2009
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===Cultures for characterization=== | ===Cultures for characterization=== | ||
IPTG : stock solution of 100 mM. Induction at 1 mM should be enough, so we have to dilute it 100x. | IPTG : stock solution of 100 mM. Induction at 1 mM should be enough, so we have to dilute it 100x. | ||
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+ | - RO2 #4 - BB1, clone #3. 4 conditions : +IPTG +light, +IPTG -light, -IPTG -light, -IPTG +light. Antibiotic : 10 ul amp + 30 ul kana. | ||
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+ | - RO1#1 -BB1, clone #1. The same 4 conditions as above. Antibio : 10 ul amp + 30 ul kana. | ||
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+ | - Negative control : BB1 (BB1.13). Antibio : 30 ul Kana. | ||
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+ | All in 5 ml M9/minimal + AA + thiamine + corresponding antibiotic. | ||
==People in the lab== | ==People in the lab== |
Revision as of 13:40, 4 October 2009
Contents |
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