IGEM:Groningen/Notebook/iGEM 2011/2011/08/26: Difference between revisions
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== | ==26-8-11== | ||
<br> | |||
<br> colony PCR of PhybB-RBS-cI-LVA-DT, PhybB-RBS-LasR-LVA-DT and PBAD-cI-LVA (3 colonies): | |||
<br> Composition master mix: | |||
<br> 10× Taq buffer: 48μl | |||
<br> dNTPs 10mM: 9.6μl | |||
<br> MgCl2: 28.8μl | |||
<br> Forward biobrickvector primer: 9.6μl | |||
<br> Reverse biobrickvector primer: 9.6μl | |||
<br> Taq polymerase: 2.4μl | |||
<br> MQ water: 372μl | |||
<br> | |||
<br> PCR conditions: | |||
<br> Pre heated lid: 111°C | |||
<br> Denaturation: 94°C for 10min | |||
<br> Cycle (33×) | |||
<br> Denaturation: 94°C for 30s | |||
<br> Annealing: 60°C for 30s | |||
<br> Extenstion: 72°C for 2min | |||
<br> Final extension: 72°C for 10min | |||
<br> Store infinite at 4°C | |||
<br> Analyse PCR samples on a 1% agarosegel | |||
<br> | |||
<br> Plasmid prep of PBAD-RBS-GFP again. | |||
<br> ND-1000 Nanodrop measurements: plot looked better than before. | |||
<br> 50 ng/microliter DNA was obtained. | |||
<br> Samples were prepared and send out for sequencing. | |||
<br> | |||
<br> Teammeeting at 15:00! | |||
<br> | |||
Revision as of 04:10, 26 August 2011
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26-8-11
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