Team:LMU-Munich/Notebook/Protocols/24 Colony PCR

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Colony PCR

Component	Volume
PCR Master Mix	2.5µl
upstream primer	0.375µl
downstream primer	0.375µl
nuclease free water to final volume of	5µl

Guidelines for Amplification by PCR

A. Denaturation:

initial: 94°C 5'

subsequent: 30

B. Annealing:

53°C 30

C. Extension

72°C 1'30

final: 8'

D. Refrigeration

16°C

E.Cycle Number

30 cycles