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Paris/July 13
From 2008.igem.org
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| + | <div style="display: inline; border: thin solid black; background: yellow">[[Paris/July 14|Tomorrow >]]</div> | ||
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Revision as of 17:57, 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
- IGEM 2008 Biobricks : use of the maximum recoverable volume minipreps
- IGEM 2007 Biobricks : use of 3µl of minipreps
