Team:Paris/August 4

PCR Screening of Ligation Transformants
Use of 8 clones of Ligation transformants n°L100 for screening PCR

Protocol of screening PCR

 * Mix


 * 50µl of Mix PCR by tube/clone
 * one toothpick of each clone's colony by tube
 * Program : Annealing 55°C - Time élongation 1'30" - Number cycle : 29

Conditions of electrophoresis

 * 10µl of ladder 1 kb
 * 20µl of screening PCR
 * migration ~30min at 100W on 0,8% gel

Results for L100
L100= D110 + D130 = RBS-tetR-ECFP-Ter

==> Conclusion : we obtain a low quantity of PCR products, but at the good size. So now, we know that we can amplify the clones that we want for L100 transformants bacteria.