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