IGEM:Groningen/Notebook/iGEM 2011/2011/08/02: Difference between revisions
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==2-8- | ==2-8-11== | ||
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<br> Colony PCR of transformants: PBAD-cI-LVA-DT, PBAD-LasR-LVA-DT, PBAD-RBS-GFP-DT, PBAD-pSB1C3 | <br> Colony PCR of transformants: PBAD-cI-LVA-DT, PBAD-LasR-LVA-DT, PBAD-RBS-GFP-DT, PBAD-pSB1C3 | ||
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<br> Checking transformants of yesterday with colony PCR: | |||
<br>Mastermix : | |||
<br> 10× Taq buffer: 78μl | |||
<br> dNTP mix 10mM: 15.6μl | |||
<br> MgCl2: 48μl | |||
<br> taq 5u/μl: 1μl | |||
<br> Forward primer BB vector 10μM: 15.6μl | |||
<br> Reverse primer BB vector 10μM: 15.6μl | |||
<br> MilliQ water: 604.5μl | |||
<br> PCR conditions: | |||
<br> 1. Pre heated lid: 111 °C | |||
<br> 2. Denaturation: 10 min. at 94°C | |||
<br> 3. Cycle 33×: | |||
<br> Denaturation: 30s at 94°C | |||
<br> Annealing: 30s at 60°C | |||
<br> Extension: 2.5 min. at 72°C | |||
<br> 4. Final extension: 10 min. at 72°C | |||
<br> 5. Store infinite at 4°C | |||
<br> Analyse on 1% agarosegel | |||
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Revision as of 07:47, 20 September 2011
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