From 2007.igem.org

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 Take bacteria in 1,5mL LB (NaCitrate 20mM; DAP 300µM) Incubation 1h at 37°C with agitation