NOMe-Seq

Active Motif's NOMe-Seq Kit is the first commercially available assay to study the relationship between multiple epigenetic modifications on the same DNA molecule. NOMe-Seq is a high resolution single-molecule Nucleosome Occupancy and Methylome sequencing kit that provides both a nucleosome footprint and a DNA methylation profile for the same DNA strand. This enables researchers to study the role of nucleosome occupancy in the context of DNA methylation for their target gene of interest. The NOMe-Seq assay works by treating fixed chromatin with a GpC methyltransferase enzyme to artificially methylate GpC dinucleotides that are not protected by nucleosomes or other proteins bound to the DNA. Due to the high abundance of GpC residues within the genome, the enzymatic treatment provides a high resolution footprint of nucleosome occupancy. Treated DNA is then subjected to bisulfite conversion and gene-specific loci are sequenced to establish the DNA methylation profile for a single DNA strand. Since GpC dinucleotides are not methylated in mammalian genomes, the endogenous CpG methylation can be distinguished from the artificially introduced GpC methylation. By combining the sequencing information for the GpC sites with the CpG sites, a profile of nucleosome occupancy and DNA methylation for a specific gene locus can be determined.