Paris/July 12

From 2007.igem.org

(Difference between revisions)
Line 8: Line 8:
** 450µl MgSO4
** 450µl MgSO4
** 90µl DAP
** 90µl DAP
-
Let grow until OD=0.2 (~1h)
+
** 150µl W121 from an ON culture
 +
* Let grow until OD=0.2 (~1h)
 +
* Add 100µl P1 (stock 10^10/ml)
 +
* Incubate 2h
 +
* Add 1ml of the ON culture of W121
 +
* Incubate 2h
 +
* Add 200µl Chloroform
 +
* Vortex, centrifuge (15min @ 3000rpm)
 +
* Recover the supernatant = Stock keep at 4°C

Revision as of 10:53, 12 July 2007

David B:

Preparation of a stock of phage on w121:

  • Filtration of the Cacl2 and MgSO4 stocks
  • Launch a culture:
    • 15ml LB
    • 225µl Cacl2
    • 450µl MgSO4
    • 90µl DAP
    • 150µl W121 from an ON culture
  • Let grow until OD=0.2 (~1h)
  • Add 100µl P1 (stock 10^10/ml)
  • Incubate 2h
  • Add 1ml of the ON culture of W121
  • Incubate 2h
  • Add 200µl Chloroform
  • Vortex, centrifuge (15min @ 3000rpm)
  • Recover the supernatant = Stock keep at 4°C