IGEM:Groningen/Notebook/iGEM 2011/2011/06/23: Difference between revisions
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Joyce Mulder (talk | contribs) (Autocreate 2011/06/23 Entry for IGEM:Groningen/Notebook/iGEM_2011) |
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== | ==23-6-11== | ||
<br> | |||
<br> PCR for PcI, PBADaraC, PlasB and sample of Christoph | |||
<br> <br> PCR with taq: | |||
<br> 10× taq buffer: 5μl | |||
<br> 10mM dNTPs: 1μl | |||
<br> BB forward primer10μM: 2.5μl | |||
<br> BB reverse primer10μM: 2.5μl | |||
<br> Taq DNA polymerase: 0.5μl | |||
<br> MgCl2 25mM: 4μl | |||
<br> MQ water: 33.5μl | |||
<br> | |||
<br> PCR with pfu: | |||
<br> 10× pfu buffer with MgSO4: 5μl | |||
<br> 10mM dNTPs: 1μl | |||
<br> BB forward primer10μM: 1μl | |||
<br> BB reverse primer10μM: 1μl | |||
<br> Pfu DNA polymerase: 1μl | |||
<br> MQ water: 40μl | |||
<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> Clean up DNA with High Pure PCR Purification Kit | |||
<br> | |||
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Revision as of 03:35, 21 September 2011
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23-6-11
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