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

Band	1-->8
Expected size	1042 pb
Measured size	1100 pb
Gel : L100(1-8)

==> 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.