Team:Macquarie Australia/PCR

pbanner

PCR



General mastermix recipe:
      4 µL of 5x phusion buffer
      0.4 µL dNTPs
      0.6 µL of DMSO
      0.2 µL of polymerase
      11.8 µL of water

To this mixture, add:
      1 µL of forward primer
      1 µL of reverse primer
      1 µL of template
      Total volume = 20 µL

    PCR settings:
        A general program for PCR is as follows:
        Initial denaturation at 98oC for 30 seconds

      Followed by 30 repeats of:
          Denaturation – 98oC for 10 seconds
          Annealing – 60oC for 10 seconds
          Extension – 72oC for 2 minutes
          Final extension – 72oC for 10 minutes