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Here we have performed the first large scale (over 300 samples) genome-wide study of DNAm profiles using a state-of-the art platform that measures the methylation state of over 27,000 CpGs.
The Illumina HumanMethylation450k microarray measures the methylation levels of more than 485,000 methylation sites.
Those investigators used an Illumina Infinium HumanMethylation27 array, which measures the methylation level at ∼27,000 CpG sites.
Our pyrosequencing assay measures the methylation levels of five adjacent CpG sites in the BRCA1 5′ promoter.
This bead chip measures the methylation status of over 27,000 CpG sites in the genome at single nucleotide resolution [ 10].
This assay measures the methylation status of seven CpG sites at positions +318, +321, +344, +351, +353, +357 and +370 relative to the sFRP1 translation start site.
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Each of these was automatically sequenced in the twelve colonies to measure the methylation status of every single CpG dinucleotide for subsequent statistical analysis.
We monitored CD25 and FOXP3 expression by flow cytometry and measured the methylation status of the FOXP3 promoter at given time points (day 0, 7, 14 and 21) using the COBRA based method previously described.
We measured the methylation status of the ALVE-region 2 in blood in the two parental lines and 6 randomly selected RCS C, F, J, L, M and T. We found that the methylation patterns can easily be categorized into two groups, as exemplified in Figure 6A and 6B.
Specifically, by measuring the methylation levels of over 27,000 CpGs in blood cells from 148 healthy individuals and 113 age-matched pre-treatment ovarian cancer cases, we derive a DNAm signature that can predict the presence of active ovarian cancer in blind test sets with an AUC of 0.8 (95% CI (0.74 0.87)).
Methylation microarrays measure the methylation levels of thousands of genes in a single assay.
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