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To exclude differences owing to variation among different bisulfite conversion protocols and reagents 70 , these samples were bisulfite-converted centrally, using the EZ DNA Methylation-Direct Kit (Zymo Research, D5020) with the following deviations from the manufacturer's protocol: Conversion reagent was applied at 0.9 concentration, reactions incubated for 20 cycles of 1 min at 95 C and 10 min at 60 C, and the desulphonation time was extended to 30 min

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[DOI] [PubMed] [Google Scholar] 145.Kharitonov SA, Donnelly LE, Montuschi P, Corradi M, Collins JV, Barnes PJ