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. Author manuscript; available in PMC: 2017 Aug 23.
Published in final edited form as: Nat Protoc. 2014 Sep 4;9(10):2285–2300. doi: 10.1038/nprot.2014.081
Cycle number Denature Anneal Extend Hold
1 95 °C, 3 min
2–26 95 °C, 15 s 58 °C, 15 s 72 °C, 15 s/kb of desired product
27 72 °C, 5 min
28 12 °C