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. Author manuscript; available in PMC: 2019 Jul 2.
Published in final edited form as: Nat Protoc. 2019 Apr 17;14(5):1661–1685. doi: 10.1038/s41596-019-0154-6
Step Denature Anneal and extend Melt curve Hold
1 95 °C, 10 min
2–41 95 °C, 15 s 60 °C, 45 s
42 60 °C→95 °C, 2 °C/s
43 25 °C