Team:Paris/August 29

=Cloning of EnvZ*=

Ligation

 * 10 min at room temperature
 * followed by 7 hours at about 16°C.

Transformation of E. coli DH5 alpha competent cells

 * Defroze of the cells on ice
 * Add 5 µL of DNA in 100 µL of competent cells (ligation products or pUC19 for positive control)
 * 30 min on ice
 * Heat schock 45sec at 42°C
 * 2 min on ice
 * Add 900 µL of SOC
 * Incubate 1h at 37°C
 * Centrifugate 5 min at 6000 rpm
 * Remove 800 µL
 * Plate the 200 µL left on LB + amplicilline
 * Incubate O/N at 37°C