IGEM:Groningen/Notebook/iGEM 2011/2011/06/10: Difference between revisions
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== | ==10-06-11== | ||
<br> | |||
<br> Colony PCR of transformants! | |||
<br> Taq 10× buffer: 40μl | |||
<br> dNTPs 10mM: 8μl | |||
<br> Forward Biobrick primer: 8μl | |||
<br> Reverse Biobrick primer: 8μl | |||
<br> Taq DNA polymerase: 2μl | |||
<br> MQ water: 334μl | |||
<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> Asked info in team about why some PCR's fail: next time use for taq buffer not the buffer with KCI, but NH4SO4 | |||
<br> | |||
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Revision as of 02:23, 21 September 2011
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