Paris/July 16
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The Lox66-DapAColi PCR did not work... We'll try again with different annealing temperature | The Lox66-DapAColi PCR did not work... We'll try again with different annealing temperature | ||
Revision as of 15:04, 16 July 2007
Contents |
Plasmid pKs::DGAT expression in E. Coli
We tried to see TG with NR died on E. Coli cells transfected with pKs::DGAT, an IPTG-inducible promoter.
We tried different growth media containing more or less oleate, that should in theory increase TG synthesis.
Results : we don't see the inductible effect of IPTG. We can think that :
- Either the fluorescence without IPTG is due to a leak of the promoter
- Either DGAT is not induce in presence of IPTG, and the fluorescence we see is only a background.
Growth kinetics of w121 strain
Results of the previous day : we lost everything because of a crash of the computer :(.
Transduction of MG1655 with P1 stock made on w121
- Control (1mL LB MgSO4 30mM; CaCl2 15mM)
- 5µL Phage + 900µL LB (MgSO4 30mM; CaCl2 15mM) + 100µL MG1655 Culture ON
- 50µL Phage + 900µL LB (MgSO4 30mM; CaCl2 15mM) + 100µL MG1655 Culture ON
- 500µL Phage + 500µL LB (MgSO4 30mM; CaCl2 15mM) + 100µL MG1655 Culture ON
=>For the nth time, it is not working : we have only contaminants.
MiniPreps
- I0500 clones 1, 2
- pJ23107 clones 1, 2
PCR purification
- Lox71-FtsZ1
- FtsZ2
- DGAT1
- DGAT2
PCRs
The Lox66-DapAColi PCR did not work... We'll try again with different annealing temperature
