Difference between revisions of "Team:UiOslo Norway/Experiments/PCR phusion"
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Revision as of 10:08, 9 September 2015
PCR with Phusion-HF DNA Polymerase (NEB):
Back to ProtocolsUsing the guidelines of the provided Phusion® High-Fidelity DNA Polymerase (M0530) protocol.
Reaction Setup: 10 µl 5X Phusion HF Buffer 5 µl dNTPS [2 mM] 2.5 µl Forward Primer [10 µM] 2.5 µl Reverse Primer [10 µM] 1 µl Template DNA 0.5 µl Phusion HF DNA Polymerase 28.5 µl MilliQ Water
50 µl Total Reaction
PCR Program:
Step Temperature Time Initial Denaturation 98 °C 30 seconds 35 Cycles 98 °C 10 seconds 45 °C – 72 °C 30 seconds 72 °C 30 seconds / 1 kb Final Extension 72 °C 5 minutes Hold 4 °C forever