Paris/July 16

From 2007.igem.org

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== Kinetics ==
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We had a problem with the computer... we lost the kinetics :-( Sorry Eimad !
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== Plasmid pKs::DGAT expression in E. Coli ==
== Plasmid pKs::DGAT expression in E. Coli ==
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* Either the fluorescence without IPTG is due to a leak of the promoter
* 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.
* Either DGAT is not induce in presence of IPTG, and the fluorescence we see is only a background.
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== Growth kinetics of w121 strain ==
== Growth kinetics of w121 strain ==
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Results of the previous day : we lost everything because of a crash of the computer :(.
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Results of the previous day : we lost everything because of a crash of the computer :( Sorry Eimad !
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== Transduction of MG1655 with P1 stock made on w121 ==
== Transduction of MG1655 with P1 stock made on w121 ==
* Control (1mL LB MgSO4 30mM; CaCl2 15mM)
* Control (1mL LB MgSO4 30mM; CaCl2 15mM)

Revision as of 15:13, 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.
Paris EColi DGAT.jpg
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 :( Sorry Eimad !

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