IGEM:Groningen/Notebook/iGEM 2011/2011/08/11: Difference between revisions
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<br> Isolate from 1% agarose gel with the agarose extraction kit | <br> Isolate from 1% agarose gel with the agarose extraction kit | ||
<br> PCR overnight for taRNA fragment: | <br> PCR overnight for taRNA fragment: | ||
<br> 10× | <br> 10× taq buffer: 5μl | ||
<br> dNTPs: 1μl | <br> dNTPs: 1μl | ||
<br> F3: 1μl | <br> F3: 1μl | ||
<br> R4: 1μl | <br> R4: 1μl | ||
<br> pfu: | <br> pfu: 0.04μl | ||
<br> taq: 0.25μl | |||
<br> DNA template: 5μl | <br> DNA template: 5μl | ||
<br> | <br> 33.71μl MQ | ||
<br> | <br> | ||
<br> PCR conditions: | <br> PCR conditions: | ||
<br> Pre heated lid: 111°C | <br> Pre heated lid: 111°C | ||
<br> Denaturation: | <br> Denaturation: 94°C for 10min | ||
<br> Cycle (35×) | <br> Cycle (35×) | ||
<br> Denaturation: | <br> Denaturation: 94°C for 30s | ||
<br> Annealing: 53°C for 30s | <br> Annealing: 53°C for 30s | ||
<br> Extenstion: 72°C for 1min | <br> Extenstion: 72°C for 1min |
Revision as of 09:15, 11 August 2011
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