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. Author manuscript; available in PMC: 2018 Jun 18.
Published in final edited form as: Nat Struct Mol Biol. 2017 Dec 18;25(1):109–114. doi: 10.1038/s41594-017-0006-4
Cycle number Denaturation Annealing Extension
0 68 °C, 10 min
1 94 °C, 30 sec 65 °C, 5 min 68 °C, 15 min
2-4 94 °C, 30 sec 65 °C, 1 min 68 °C, 10 min
5-21 94 °C, 30 sec 65 °C, 1 min 68 °C, 2 min
22 68 °C, 5 min