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. Author manuscript; available in PMC: 2016 Jul 1.
Published in final edited form as: Nat Protoc. 2015 Jun 11;10(7):985–1006. doi: 10.1038/nprot.2015.065
Cycle number Denature Anneal Extend Final
1 95 °C, 10 min
2–62 95 °C, 10 s 58 °C, 45s 72 °C, 30 s
Melting curve 95 °C, 60s 55 °C, 60s followed by 55 + 0.5 °C, 10s each
64
65
Hold 4 °C