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
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