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. Author manuscript; available in PMC: 2018 Nov 1.
Published in final edited form as: Nat Protoc. 2018 Apr 5;13(5):875–898. doi: 10.1038/nprot.2018.007
Cycle number Denature Anneal Extend
1 95 °C, 3 min
2–32 95 °C, 20 s 57 °C, 20 s 72 °C, 60 s per kb
33 72 °C, 5 min