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. Author manuscript; available in PMC: 2013 Aug 9.
Published in final edited form as: Nat Protoc. 2011 Mar 24;6(4):482–501. doi: 10.1038/nprot.2011.301
Cycle number Denature Anneal Extend
1 95 °C, 2 min — —
2–35 95 °C, 30 s 5 °C below Tm, 30 s 72 °C, 1 min per kb
36 — — 72 °C, 10 min