Paris/July 13

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(Difference between revisions)
(Transformation of Biobricks from the iGEM2007 plates)
(Transformation of Biobricks from the iGEM2007 plates)
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== Transformation of Biobricks from the iGEM2007 plates ==
== Transformation of Biobricks from the iGEM2007 plates ==
-
* BBa_I0500: Inducible pBad/araC  
+
* BBa_I0500: Inducible pBad/araC in  pSB2K3
-
well: 9I,  Plate: pSB2K3
+
well: 9I,  Plate 2   
-
*
+
 
 +
* BBa_J23100: strongest constitutive promoter in BBa_J61002
 +
well: 21E, Plate 3
 +
 
 +
 
 +
 
 +
* BBa_B0015: double terminator (B0010-B0012) in pSB1AK3
 +
well: 1I, Plate 1

Revision as of 12:27, 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

well: 9I, Plate 2

  • BBa_J23100: strongest constitutive promoter in BBa_J61002

well: 21E, Plate 3


  • BBa_B0015: double terminator (B0010-B0012) in pSB1AK3

well: 1I, Plate 1