Lidstrom:Colony PCR

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Revision as of 15:11, 2 February 2012 by Janet B. Matsen (talk | contribs) (New page: ==General:== *The thermocycling denaturation & extension temperature should be that recommended by your particular PCR kit. I have seen 68oC & 72oC. *The annealing temperature is up to y...)
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General:

  • The thermocycling denaturation & extension temperature should be that recommended by your particular PCR kit. I have seen 68oC & 72oC.
  • The annealing temperature is up to you. For VF2 & VR 54-56oC works well.
  • The primer concentration varies in different people's recipes.
  • Janet is using NEB's OneTaq quick-load (not hot start) & their recommended protocol