Team:Freiburg/Protocols/PCR

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PCR

Programm normale PCR

Lid temperature: 98°C

98°C	5 min	Denaturierung
98°C	30 s	Denaturierung
63°C	30 s	Annealing
72°C	30 - 40 s pro kb	Elongation
72°C	10 min	Restelongation
4°C	hold

18 - 25 Zyklen (Schritt 2-4)

PCR mix

ingredient	volume
Template	200 - 300 ng
Primer (10 µM)	1 µl each
Q5 Polymerase	0.5 µl
dNTPs	4 µl
DMSO	1 µl
Q5 reaction buffer (5x)	10 µl
dH2O	up to 50 µl

Add GC enhancer if the GC content of the template [??] is very high.

protocols old