Raw data (beta values) was extracted from Illumina Bead Studio (version V2011.1). Methylation data was further processed via the Bioconductor lumi package [51 (link)], which works on M values rather than on beta values. The relationship between beta and M values is given as [52 (link)]:
We used shift-scale color bias adjustment and quantile normalization as further preprocessing steps implemented in the lumi package. A detection p value cutoff of 0.00001 was used to filter out signals below background, and only CpGs in autosomes were considered. We used limma [53 ] to assess differential methylation of CpG sites between patient groups. Cross reactive loci were defined as described by Chen Y et al, [54 (link)]