Paris/July 13
From 2007.igem.org
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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. | ||
| - | + | <br> | |
| - | First step for transduction : adsorption | + | First step for transduction : adsorption<br> |
| - | use of 03/07/07 (I) and 12/07/07 (II) phage stock from W121 : X2 | + | use of 03/07/07 (I) and 12/07/07 (II) phage stock from W121 : X2<br> |
| - | MG1655 culture ON | + | MG1655 culture ON<br> |
| - | + | * 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 | |
| - | + | <br> | |
| - | 20 min at 37°C | + | 20 min at 37°C<br> |
| - | Centrifuge at 12000rpm during 5 min | + | Centrifuge at 12000rpm during 5 min<br> |
| - | Take bacteria in 1,5mL LB (NaCitrate 20mM; DAP 300µM) | + | Take bacteria in 1,5mL LB (NaCitrate 20mM; DAP 300µM)<br> |
| - | Incubation 1h at 37°C with agitation | + | Incubation 1h at 37°C with agitation<br> |
Revision as of 10:15, 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
