Team:Paris/September 5

=Purification of yesterday digestion=



Electrophoresis and gel extraction
1% agarose gel

Qiaquick Gel Extraction Kit
 * elution in 30 µL of EB buffer

Concentration measurement
=Constructions=

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