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Paris/July 13
From 2008.igem.org
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[[Paris/July 12|< Yesterday]] - [[Paris/July 14|Tomorrow >]] | [[Paris/July 12|< Yesterday]] - [[Paris/July 14|Tomorrow >]] | ||
| - | == Preparation of Competent Cells == | + | [[== Preparation of Competent Cells ==]] |
| - | * Dilution 1/200e (1mL of culture on 200mL of LB) | + | * Dilution 1/200e (1mL of the O/N culture on 200mL of LB) |
* Culture at 37°C until DO=0,57 (around 3h) | * 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 | * Sharing into 4 Falcon tubes of 50mL and cooling at 4°C during 40' in ice | ||
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* Spin during 8', 2000rpm, 4°C | * Spin during 8', 2000rpm, 4°C | ||
* Resuspension of the cell's pellet in 1mL of cold CaCl2 (0,1M) | * Resuspension of the cell's pellet in 1mL of cold CaCl2 (0,1M) | ||
| - | * 1h of delay | + | * 1h of delay unexpected (problem with DNA lyophilized) |
| + | * Transformation of the Biobricks with the "classical" protocol (--> heat shock at 42°C durin 40") | ||
| + | * Incubation O/N at 37°C of the transformed bacteria | ||
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| + | |||
| + | [[Remark :]] | ||
| + | IGEM 2008 Biobricks : use of the maximum recoverable volume minipreps | ||
| + | IGEM 2007 Biobricks : use of 3µl of minipreps | ||
Revision as of 17:39, 15 July 2008
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== Preparation of Competent Cells ==
- Dilution 1/200e (1mL of the O/N 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 unexpected (problem with DNA lyophilized)
- Transformation of the Biobricks with the "classical" protocol (--> heat shock at 42°C durin 40")
- Incubation O/N at 37°C of the transformed bacteria
Remark :
IGEM 2008 Biobricks : use of the maximum recoverable volume minipreps
IGEM 2007 Biobricks : use of 3µl of minipreps
