Paris/July 12
From 2007.igem.org
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David.bikard (Talk | contribs) |
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** 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
