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. Author manuscript; available in PMC: 2020 May 26.
Published in final edited form as: Nat Protoc. 2017 Feb 16;12(3):581–603. doi: 10.1038/nprot.2016.165
Cycle number Denature Anneal Extend
1 95 °C, 15 min
2–16 95 °C, 30 s *72 °C, 30 s 72 °C, 30 s
17–40 95 °C, 30 s 58 °C, 30 s 72 °C, 30 s
41 72 °C, 20 min
*

Decrease annealing temperature by 1 °C in each subsequent cycle.