Five DNA samples chosen at random from cases and five from controls were analyzed using the cloning method. The methylation pattern of each is shown in Figure 2A. We also analyzed, with this method, five selected samples of cases previously evaluated by sequencing/ESME analysis (Figure 2B). One had the highest percent methylation at the −25 CpG site (1, Figure 2B), two had the highest methylation at the +84 CpG site (2 and 3), one had intermediate levels of methylation across the CpG sites (4), and the final sample (5) had low levels of methylation across the sites. All 15 samples were analyzed by sequencing/ESME analysis. Twelve samples had a forward to reverse correlation ≥ 0.7; for these, the correlation of the percentage of methylation was R2 =0.79 (Figure 2C).
