Paris/July 13
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== Transduction of MG1655 with P1 stock made on w121 == | == 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. | 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 | ||
Revision as of 10:13, 13 July 2007
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
