Lidstrom:Colony PCR

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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