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. Author manuscript; available in PMC: 2021 Oct 19.
Published in final edited form as: Nat Protoc. 2021 Feb 22;16(3):1647–1713. doi: 10.1038/s41596-020-00478-x
Cycle no. Denature Anneal Extend no. 1 Imagea Extend no. 2
1 98 °C, 3 min - - - -
2–32 98 °C, 5 s 65 °C, 10 s 72 °C, 10 s Image 72 °C, 8 s
a
This is the step where the quantitative PCR machine images each well to see how much double-stranded DNA (dsDNA) is detected by EvaGreen Dye labeling.