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. Author manuscript; available in PMC: 2018 Apr 3.
Published in final edited form as: Curr Protoc Mol Biol. 2017 Apr 3;118:15.15.1–15.15.15. doi: 10.1002/cpmb.38
Initial denaturation: Denature: 5 min 95°C
6 cycles: Denature: 30 sec 95°C
Anneal: 30 sec 30°C
Extend: 60 sec 72°C
30 cycles: Denature: 30 sec 95°C
Anneal: 30 sec 45°C
Extend: 120 sec 72°C
Final extension: Extend: 4 min 72°C
Hold: ∞ 12°C