IGEM:Groningen/Notebook/iGEM 2011/2011/08/02

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2-8-11



Colony PCR of transformants: PBAD-cI-LVA-DT, PBAD-LasR-LVA-DT, PBAD-RBS-GFP-DT, PBAD-pSB1C3

Checking transformants of yesterday with colony PCR:
Mastermix :
10× Taq buffer: 78μl
dNTP mix 10mM: 15.6μl
MgCl2: 48μl
taq 5u/μl: 1μl
Forward primer BB vector 10μM: 15.6μl
Reverse primer BB vector 10μM: 15.6μl
MilliQ water: 604.5μl
PCR conditions:
1. Pre heated lid: 111 °C
2. Denaturation: 10 min. at 94°C
3. Cycle 33×:
Denaturation: 30s at 94°C
Annealing: 30s at 60°C
Extension: 2.5 min. at 72°C
4. Final extension: 10 min. at 72°C
5. Store infinite at 4°C
Analyse on 1% agarosegel