Team:Paris/July 16

Results of transformations

 * Positive controls from biobricks 2007, highest number of bacterias than the first try. It's probably due to the highest quantity of bacteria introduced (150µL instead 100µL).


 * Transformations from biobricks 2008 don't succeed again.

Preparation of competent cells

 * Dilutions 1/200
 * Culture during 3h --> DO= 0,15 /  4h --> DO= 0,13
 * problem from the spectro, culture have been thrown.

Transformations Biobricks 2008
use of a new protocol --> directly competent cells


 * Defroze competent cells on ice during 5'
 * Mix 100µL of competent bacterias with 5µL of biobricks 08 (or 1µL for the positive control)
 * Incubate 10' on ice
 * Heat-shock the cells during 45-50" at 42°C (don't shake!!!)
 * Put 2' on ice
 * Add 900µL of cold SOC medium (4°C)
 * Incubate 1h at 37°C under shaking (225rpm)
 * Transfer in an eppendorf tube and spin at 5000rpm during 30"
 * Remove 800µL of supernatant
 * Resuspent the pellet in the 150-200µL left
 * Spread on plates
 * Incubate O/N at 37°C