User:Tim Henry/Notebook/D1S80/2012/02/24

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D1S80 PCR

Looking at datasheet for Maxime i-Taq, it suggest using 10pM of primer.

From primer datasheet it appears it is 200mL, 100μM - if I have this correct it should be 0.5μM/1μL. This would correspond to protocols for other taq polymerases (0.1-1μM suggested).

To dilute, 49μL dH2O + 1μL. Then use 1μL (10pM)

1.5 μL template (usually use 2.0)

1μL primer each

16.5μL dH2O

30 cycles of denature/anneal/extension

  • 94°C 3min initial
  • 94°C 30s denature
  • 67°C 30s anneal
  • 72°C 30s extension
  • 72°C ~5-7min (auto to next program)

Put template + primers on 94°C first before adding taq, pseudo hot start style.