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. Author manuscript; available in PMC: 2015 May 1.
Published in final edited form as: Nat Protoc. 2014 Oct 9;9(11):2586–2606. doi: 10.1038/nprot.2014.170
Cycle Denature Anneal Extend
1 95 °C for 4 min
2-Variable 98 °C for 20 s 60 °C for 20 s 72 °C for 15 s