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. Author manuscript; available in PMC: 2015 Jan 1.
Published in final edited form as: Nat Protoc. 2014 Jun 12;9(7):1607–1620. doi: 10.1038/nprot.2014.105
Cycle Number Denature Anneal Extend Hold
1 98°C, 10 sec 9
2-6 98°C, 15 sec 64°C-1°C/cycle, 5 sec 68°C, 1 min/kb + 1 min
7-27 98°C, 15 sec 60°C, 25 sec 68°C, 1 min/kb + 1 min
28 4°C