IGEM:Groningen/Notebook/iGEM 2011/2011/05/18

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18-5-11



2PCRs failed yesterday, byproducts
Try to resuspend them better and leave them on ice after pipetting


Searching for protocols
Do a PCR for PhybB on E.coli DH5alpha suspension:
Composition:
Taq 10× buffer: 5μl
dNTPs: 1μl
MgCl2: 3μl
taq 5u/μl: 0.25μl
Forward primer 10mM: 1μl
Reverseprimer 10mM: 1μl
Template: 1μl
MQ: 38.75μl

Analyse them on a 1% agarosegel
PCR conditions:
denaturation: 10min
Cycle 25×
denaturtaion: 1min
annealing: 1min (gradient for 60, 65 and 70 degrees)
extension: 1min
Final extension: 10min
Store infinite


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