Paris/July 13
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== Transformation of Biobricks from the iGEM2007 plates == | == Transformation of Biobricks from the iGEM2007 plates == | ||
| - | * BBa_I0500: Inducible pBad/araC in pSB2K3 | + | * BBa_I0500: Inducible pBad/araC in pSB2K3 (KanR) |
well: 9I, Plate 2 | well: 9I, Plate 2 | ||
| - | * BBa_J23100: | + | * BBa_J23100: strong constitutive promoter in BBa_J61002 (AmpR) |
well: 21E, Plate 3 | well: 21E, Plate 3 | ||
| - | + | * BBa_B0015: double terminator (B0010-B0012) in pSB1AK3 (AmpR) | |
| - | + | ||
| - | * BBa_B0015: double terminator (B0010-B0012) in pSB1AK3 | + | |
well: 1I, Plate 1 | well: 1I, Plate 1 | ||
Revision as of 12:41, 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)
Transformation of Biobricks from the iGEM2007 plates
- BBa_I0500: Inducible pBad/araC in pSB2K3 (KanR)
well: 9I, Plate 2
- BBa_J23100: strong constitutive promoter in BBa_J61002 (AmpR)
well: 21E, Plate 3
- BBa_B0015: double terminator (B0010-B0012) in pSB1AK3 (AmpR)
well: 1I, Plate 1
