Team:Washington/Protocols/PCR
From 2011.igem.org
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Revision as of 22:14, 13 September 2011
General PCR Protocol
- Setup Amplification PCR Reaction
- 1uL Template
- 1uL 25mM dNTP's
- 10uL Phusion HF Buffer
- 0.5uL Forward Primer (Tm 65)
- 0.5uL Reverse Primer (Tm 65)
- 0.5uL Phusion polymerase
- 36.5uL diH2O
- Amplification PCR Reaction
- 98C - 30s
- 98C - 10s
- 63C - 10s
- 72C - 30s/kb target gene
- Repeat 2-4 29x
- 72C - 5min
- 10C - forever
Refer to NEB's online protocol for Phusion http://www.neb.com/nebecomm/products/protocol87.asp