Team:Paris Bettencourt/Protocols/PCR

PCR protocol (Phusion)
  • Prepare the following mix
    • 2µL primer Forward (10 µM)
    • 2µL primer Reverse (10 µM)
    • 0.1 to 1 ng of template DNA
    • 25µL of Life Technologies Phusion High-Fidelity PCR Master Mix with HF Buffer
    • complete to 50µL of DNase/RNase-free water
  • Launch 30 to 35 PCR cycles using the following parameters:
    time (min) temperature (°C) function
    3:00 98 melting
    0:30 98 melting
    0:30 XX°C annealing
    1 minute/kb 72 extension
    10:00 72 extension
    infinite hold 12 storage