Difference between revisions of "Team:UiOslo Norway/Experiments/PCR phusion"

 
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Latest revision as of 12:09, 18 September 2015

PCR with Phusion-HF DNA Polymerase (NEB):

Back to Protocols


Using the guidelines of the provided Phusion® High-Fidelity DNA Polymerase (M0530) protocol.

  1. 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


  2. 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

    3. iGEM UiOslo 2015 is sponsored by: