Paris/July 13

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Revision as of 11:28, 13 July 2007 by Landrain (Talk | contribs)

Transduction of MG1655 with P1 stock made on w121

The isolated clones of FtsZ TS all grew at 42°C. They must be mutants... We'll do the transduction only on MG1655 for the moment.
First step for transduction : adsorption
use of 03/07/07 (I) and 12/07/07 (II) phage stock from W121 : X2
MG1655 culture ON

  • 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


20 min at 37°C
Centrifuge at 12000rpm during 5 min
Resuspend bacteria in 1,5mL LB (NaCitrate 20mM; DAP 300µM)
Incubation 1h10' at 37°C with agitation
Centrifuge 5 min at 5000rpm

Plate bacteria on LB-agar (NaCitrate 20mM; DAP 300µM)

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