Team:Paris/September 3

=Construction of rbs-TetR-mRFP-LVA-tripart (L173)=

Transformation
Ligation L173: Insert:vector ratio : 3:1 or 4:1
 * insert: D112 (ES) rbs-TetR
 * vector: D187 (EX) mRFP-LVA-tripart

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