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Table 1 BS conversion protocols and parameters

From: Comparison of whole-genome bisulfite sequencing library preparation strategies identifies sources of biases affecting DNA methylation data

Method Denaturation temperature Conversion temperature Incubation time
Heat 1 High heat (99 °C) 65 °C 90 minutesa
Heat 2 High heat (99 °C) 55 °C 10 hb
Alkaline 1 Low heat (37 °C) 65 °C 90 minutesa
Alkaline 2 Low heat (37 °C) 50 °C 12–16 hc
Am-BS Low heat (37 °C) 70 °C 30 minutesd
  1. aImprint DNA Modification Kit (Sigma-Aldrich), one-step protocol for ‘Heat 1’ and 2-step protocol for ‘Alkaline 1’
  2. bEpiTect Bisulfite kit (Qiagen), FFPE protocol and doubled incubation time (see “Methods”)
  3. cEZ DNA Methylation Kit (Zymo Research)
  4. dProtocol conditions as described in Hayatsu et al. [25]