Methylome Analysis

DNA methylation is the most studied form of epigenetic modification of the genome. Thought primarily to be related to the regulation of gene expression via the inhibition of transcription factor binding, analysis of methylated cytosines within a genome, and across a species’ genomes, can shed light on which genes and regulatory elements are required, or not, at a given point in time. Therefore, the methylome – a genome’s collection of methyl marks – is a changeable snapshot of an organism’s (or a population’s) genomic response to its environment.

Historically, whole genome bisulfite sequencing (WGBS) has been the standard technique for methylome sequencing. However, WGBS relies on harsh chemical treatments at high temperatures that are extremely damaging to DNA, resulting in fragmentation, loss and bias. These challenges with bisulfite sequencing led to the development of an enzyme-based alternative (NEBNext^® Enzymatic Methyl-seq, or EM-seq™) that minimizes DNA damage and provides superior detection of 5mC and 5hmC from fewer sequencing reads. EM-seq has now been further enhanced with the NEBNext Enzymatic Methyl-seq v2 Kit which accommodates an expanded input range (as low as 100 pg), a more streamlined, faster workflow, and improved performance. The NEBNext Enzymatic 5hmC-seq Kit (E5hmC-seq™) further expands the product line, enabling specific detection of 5hmC.

This animated workflow describes the more-gentle NEBNext EM-seq alternative to bisulfite-based conversion.

EM-seq™ conversion method

The EM-seq™v2 workflow accommodates a wider input range than the original EM-seq workflow, with a 100-fold lower minimum input amount. The v2 workflow is more streamlined, has one fewer cleanup step and is 30 minutes faster. Note that NEBNext LV UDI primers are not included in the kit and are available separately.

The EpiMark^® suite of products were the first generation of New England Biolabs’ epigenetics offering. You can find the current list of available EpiMark products here.