Alm:PCR

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Revision as of 12:55, 21 September 2007 by ClarkeS (Talk | contribs)
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Standard PCR reactions

  • ~100 ng template
  • ~100 pmole each primer
  • buffer to 1X
  • polymerase
  • denature 94-95°C
  • anneal 5°C less than lowest primer hyb temp
  • extend 1’/kb to be amplified

PCR Master Mix (2.5X)

  • 62.5 U/ml Taq DNA Polymerase
  • 125 mM KCl
  • 75 mM Tris-HCl, pH 8.3
  • 3.75 mM Mg(OAc)2
  • 500 uM each dNTP


5'-CATTAG can be useful to add to prevent exonuclease activity from degrading recognition sites

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