Paris/July 13
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20 min at 37°C<br> | 20 min at 37°C<br> | ||
Centrifuge at 12000rpm during 5 min<br> | Centrifuge at 12000rpm during 5 min<br> | ||
| - | + | Resuspend bacteria in 1,5mL LB (NaCitrate 20mM; DAP 300µM)<br> | |
| - | Incubation | + | Incubation 1h10' at 37°C with agitation<br> |
| + | Centrifuge 5 min at 5000rpm<br> | ||
| + | <br> | ||
| + | Plate bacteria on LB-agar (NaCitrate 20mM; DAP 300µM)<br> | ||
Revision as of 11:28, 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
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)
