Team:Heidelberg/Templates/Methods

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Colony PCR

Colonies were picked with pipet tips and dipped into the PCR mixture.

Ingredient amount [µl]
Overnight Culture Top of a pipet tip
10x PCR buffer 2.0
MgCl2 (25mM) 2.0
Forward primer (10µM) 0.2
Reverse primer (10µM) 0.2
dNTPs (10mM) 1.0
Polymerase 0.5
Sterile water ad 10.0
Total 10.0

The following PCR program was adjusted according to annealing conditions.

Cycles temperature [°C] Time [min:s]
1 98 2:00
35 98 0:05
68 (annealing) 0:15
72 1:00 per kbp
1 72 10:00
1 12 inf