IGEM:Groningen/Notebook/iGEM 2011/2011/05/18: Difference between revisions
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Joyce Mulder (talk | contribs) (Autocreate 2011/05/18 Entry for IGEM:Groningen/Notebook/iGEM_2011) |
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== | ==18-5-11== | ||
<br> | |||
<br> 2PCRs failed yesterday, byproducts | |||
<br> Try to resuspend them better and leave them on ice after pipetting | |||
<br> | |||
<br> | |||
<br> Searching for protocols | |||
<br> Do a PCR for PhybB on E.coli DH5alpha suspension: | |||
<br> Composition: | |||
<br> Taq 10× buffer: 5μl | |||
<br> dNTPs: 1μl | |||
<br> MgCl2: 3μl | |||
<br> taq 5u/μl: 0.25μl | |||
<br> Forward primer 10mM: 1μl | |||
<br> Reverseprimer 10mM: 1μl | |||
<br> Template: 1μl | |||
<br> MQ: 38.75μl | |||
<br> | |||
<br> Analyse them on a 1% agarosegel | |||
<br> PCR conditions: | |||
<br> denaturation: 10min | |||
<br> Cycle 25× | |||
<br> denaturtaion: 1min | |||
<br> annealing: 1min (gradient for 60, 65 and 70 degrees) | |||
<br> extension: 1min | |||
<br> Final extension: 10min | |||
<br> Store infinite | |||
Revision as of 02:45, 1 August 2011
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18-5-11
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