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. Author manuscript; available in PMC: 2017 Aug 8.
Published in final edited form as: Nat Protoc. 2016 Sep 29;11(11):2104–2121. doi: 10.1038/nprot.2016.126
Cycle number Denature Anneal Extend
1 95°C, 3 min.
2–6 98°C, 20 sec. 60°C, 20 sec. 72°C, 1 min.
7–17* 98°C, 20 sec. 65°C, 20 sec. 72°C, 1 min.
*
Use optimized cycle number