Quantification of locus-specific epigenetic modifications in the mammalian genome is an integral part of understanding epigenomic changes during development and disease. The EpiMark 5-hmC and 5-mC Analysis Kit from New England Biolabs offers a simple, reproducible, and robust method to examine the methylation status of a DNA population; addresses the need for detection and quantitation of 5-hmC in genomic DNAs; and opens up opportunities to determine whether 5-hmC modifications of DNA can be exploited in the identification of novel biomarkers.
Further, quantitative PCR can be replaced by end-point PCR, providing scientists with a simple “yes or no” answer to whether a given loci contains 5-hmC. Lastly, multiplexing of this protocol for end-point PCR may be performed for different amplicons using standard thermocyclers.
The EpiMark method followed by qPCR analysis is currently being applied to study the differences in hydroxymethylation patterns for a variety of biologically relevant modifications in mammalian DNAs during the different stages of development. In unpublished studies, researchers at NEB have shown variations in 5-hmC levels of genomic DNAs extracted from brain, heart and liver tissue samples from murine and human.