Team:Paris/July 13

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 during 40")
 * Incubation O/N at 37°C of the transformed bacteria

Remark

 * Biobricks 2007 : use of 3µl of the minipreps
 * Biobricks 2008 : use of the maximum recoverable volume of the minipreps

List of the transformations

 * Biobricks 2007


 * Biobricks 2008

Stock solution

 * TE (10:1) pH 8.0
 * plates of Ampicilin and Kanamycin