Paris/July 13
From 2008.igem.org
(Difference between revisions)
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* Sharing into 4 Falcon tubes of 50mL and cooling at 4°C during 40' in ice | * Sharing into 4 Falcon tubes of 50mL and cooling at 4°C during 40' in ice | ||
* Spin during 8', 2000rpm, 4°C | * Spin during 8', 2000rpm, 4°C | ||
- | * Resuspension of the cell's | + | * Resuspension of the cell's pellet in 20mL of cold CaCl2 (0,1M) during 30' in ice |
+ | * Spin during 8', 2000rpm, 4°C | ||
+ | * Resuspension of the cell's pellet in 1mL of cold CaCl2 (0,1M) | ||
+ | * 1h of delay unexcepted (problem with lyophilized DNA) |
Revision as of 17:31, 15 July 2008
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Preparation of Competent Cells
- Dilution 1/200e (1mL of culture on 200mL of LB)
- Culture at 37°C until DO=0,57 (around 3h)
- Sharing into 4 Falcon tubes of 50mL and cooling at 4°C during 40' in ice
- Spin during 8', 2000rpm, 4°C
- Resuspension of the cell's pellet in 20mL of cold CaCl2 (0,1M) during 30' in ice
- Spin during 8', 2000rpm, 4°C
- Resuspension of the cell's pellet in 1mL of cold CaCl2 (0,1M)
- 1h of delay unexcepted (problem with lyophilized DNA)