primers with the T7 promoter tag on the reverse primer. The in vitro transcription was done, followed by site-specific cleavage using MassCLEAVE biochemistry. MassARRAY compact MALDI-TOF mass was used to acquire spectral peaks and converted into methylation ratios using EpiTYPER software72 (Ehrich et al., 2005). Two statistical criteria were used to identify genes validated by Sequenom; one with genes showing a methylation difference of greater than 10% in at least one CpG unit; furthermore, another criteria was the methylation change in the same direction of all CpG units tested for that gene amplicon (Liu et al. 2009).
