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

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Colony PCR

Component	Volume
PCR Master Mix	2.5µl
primer forward (BioBrick)	0.375µl
primer revers (BioBrick)	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