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. Author manuscript; available in PMC: 2013 May 2.
Published in final edited form as: Nat Protoc. 2012 Nov 29;7(12):2159–2170. doi: 10.1038/nprot.2012.137

Cycle number Denature Anneal Extend

1 95 °C, 2 min
2–41 95 °C, 30 s 57 °C, 30 s for 5-mC test 72 °C, 1 min
45 °C, 30 s for 5-hmC test
42 72 °C, 10 min