Phusion PCR

GroseLab:Protocols

=PCR Protocol for Phusion Polymerase=

Reference: Adapted from New England BioLabs Phusion Hot Start High-Fidelity DNA Polymerase documentation.


 * For difficult templates replace HF Buffer with GC Buffer and add DMSO or Betaine as required.


 * Do not exceed 2 U polymerase/50 ul (0.04 U/ul), especially for amplicons > 5 kb.

98 x 0:30, x (98 x 0:10, ___ x 0:20, 72 x ___), 72 x 7:00, 4 x Infinity

Primers and Tms

Forward:

Reverse:

Amplicons

=TM Calculator for Phusion=