Team:Arizona State/Lab/Protocols/PCR

From 2011.igem.org


Protocols: PCR


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  1. For 50 ul reactions, prepare master mix as follows for n tubes:
    10*n uL 5x Phusion buffer
    4*n uL 2.5 mM dNTPs
    2.5*n uL forward primer
    2.5*n uL reverse primer
    29.5*n uL PCR water
  2. Add 48 uL master mix to each of seven tubes.
  3. Label tubes blank, 0, 1, 2, 4, 8, 10
  4. Add DNA and MgCl2 to tubes according to chart:
blank 0 1 2 4 8 10
MgCl2 0 0 .5 1 2 4 5
DNA 0 1 1 1 1 1 1
  1. Add 0.5 uL Phusion polymerase to each tube.
  2. Place tubes in thermocycler.
  3. Adjust settings as needed.