The EpiMark Methylated DNA Enrichment Kit (NEB #E2600) can be used to enrich double-stranded methyl-CpG from fragmented genomic DNA with as little as 5 ng of input DNA. The EpiMark® Methylated DNA Enrichment Kit utilizes the methyl-CpG binding domain of human MBD2a protein as a capture agent. The protein is fused to the Fc tail of human IgG1 (MBD2a-Fc), which is coupled to Protein A Magnetic Beads (MBD2a-Fc/Protein A Bead). This stable complex will selectively bind double-stranded methylated CpG containing DNA. The high binding affinity of the beads coupled with optimized reagents increases sensitivity and accuracy. This kit contains all the individual components necessary to achieve enrichment in less than two hours using a four step process.
® Bisulfite Conversion Kit (NEB #E3318) is designed for the detection of methylated cytosine, using a series of alternating cycles of thermal denaturation, followed by incubation with sodium bisulfite. This kit includes all the reagents necessary for complete bisulfite conversion, including spin columns. Amplification of bisulfite-treated samples can then be performed using EpiMark® Hot Start Taq DNA Polymerase (NEB #M0490).
Mammalian DNA is enzymatically modified at the 5th carbon position of cytosine (C) bases to 5-mC, predominantly in the context of CpG dinucleotides. 5-mC is amenable to enzymatic oxidation to 5-hmC by the Tet family of enzymes, which are believed to be involved in development and disease. Currently, the biological role of 5-hmC is not fully understood, but is generating a lot of interest due to its potential as a biomarker. Common research techniques, including bisulfite sequencing methods, are unable to easily distinguish between 5-mC and 5-hmC. The EpiMark® 5-hmC and 5-mC Analysis Kit (NEB #E3317) provides a solution for distinguishing between these two modifications at specific loci.
The EpiMark® Nucleosome Assembly Kit (NEB #E5350) is used to make unmodified recombinant human nucleosomes with user-supplied DNA or the provided control DNA, and can be used for ChIP and HAT assays, as well as for enzymatic modification assays (e.g., methylation).
NEB also offers the NEBNext® ChIP-Seq Library Prep series of Master Mixes and Reagent sets that are ideally suited for sample preparation for next-generation sequencing. Each of these components must pass rigorous quality control standards and are lot controlled, both individually and as a set of reagents.
EpiMark® and NEBNext® are registered trademarks of New England Biolabs, Inc.