Quantitative Region-Specific DNA Methylation Analysis by the EpiTYPER™ Technology.

DNA methylation plays a profound role in development and health as well as development and progression of disease. High-throughput quantitative DNA methylation analysis is therefore crucial for the study of the normal physiology of the epigenome and its dysregulation in disease. Many target areas are identified by a range of emerging genome-wide cytosine methylation techniques, but these whole genome scans usually only provide methylation data for a few individual CpG sites (CpGs) within a region. The EpiTYPER™ assay is a region-specific method for the detection and quantitative analysis of DNA methylation that allows performing a high-resolution scan of selected regions. It thus enables a more detailed analysis of single CpGs and the surrounding area and can, in addition to candidate gene methylation analysis, be used to validate CpGs detected by genome wide techniques. The EpiTYPER™ assay allows a fast and reproducible targeted quantification of individual CpGs in a high throughput manner and is based on base-specific cleavage of bisulfite-converted genomic DNA and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Up to 85% of the CpGs within a target region can be analyzed and the detection precision allows quantifying methylation differences as low as 5-7%.