IGEM:Groningen/Notebook/iGEM 2011/2011/06/03
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| + | <br> Colony PCR of transformants | ||
| + | <br> Mastermix: | ||
| + | <br> Taq 10× buffer: 116μl | ||
| + | <br> dNTPs 10mM: 23.2μl | ||
| + | <br> MgCl2: 69.6μl | ||
| + | <br> Forward Biobrick primer: 23.2μl | ||
| + | <br> Reverse Biobrick primer: 23.2μl | ||
| + | <br> Taq DNA polymerase: 5.8μl | ||
| + | <br> MQ water: 899μl | ||
| + | <br> | ||
| + | <br> PCR conditions: | ||
| + | <br> Preheated lid: 111°C | ||
| + | <br> Denaturation: 94°C for 10 min. | ||
| + | <br> Cycle 33×: | ||
| + | <br> denaturation: 94°C for 30s. | ||
| + | <br> annealing: 60°C for 30s. | ||
| + | <br> Extension: 72°C for 2,5 min. | ||
| + | <br> Final extension: 72°C for 10 min. | ||
| + | <br> Store infinite at 4°C | ||
| + | <br> | ||
| + | <br> Analyse on a 1% TBE agarose gel. | ||
| + | <br> | ||
| + | <br> Working on Human Practices. | ||
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