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.
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.
Choose Type:
- Is it normal if the FE(II) solution from the EM-seq product is yellow or a color change is observed?
- Can the EM-seq Adaptor be substituted with another adaptor?
- Can the EM-seq Adaptor be used for bisulfite sequencing?
- What is the expected size of an EM-seq library?
- Can enzymatically fragmented DNA be used as input material for EM-seq™?
- Can EM-seq libraries be prepared from FFPE DNA?
- How should EM-seq libraries be sequenced?
- What is the concentration of the EM-seq Adaptor and EM-seq Index Primers?
- Are EM-seq libraries directional or non-directional?
- How should EM-seq sequencing data be analyzed?
- What is the concentration of the EM-seq Adaptor and Index Primers?
- What levels of conversion are typical with the control DNAs supplied in the EM-seq kit?
- Can other buffers be used in place of the supplied EM-seq Elution Buffer?
- How are EM-seq libraries prepared from cell-free DNA (cfDNA)?
- Why, at some stages of the EM-seq protocol, do the NEBNext Sample Purification Beads behave differently when cleaning up the sample?
- Are Sample Sheets available for use with the NEBNext® Multiplex Oligos for EM-seq?
- Can I use the NEBNext Ultra II FS DNA PCR-free Library Prep Kit for bisulfite conversion or EM-seq™ workflows?
- What is the difference between the NEBNext Enzymatic Methyl-seq Kit and the Enzymatic Methyl-seq Conversion Module?
- What is provided in the NEBNext Multiplex Oligos for Enzymatic Methyl-seq?
- Do I need to spike in custom sequencing primers when sequencing libraries made with NEBNext® Multiplex Oligos for Enzymatic Methyl-seq on Illumina® sequencing instruments?
- Are the NEBNext® Multiplex Oligos for Enzymatic Methyl-seq the same as NEBNext® Multiplex Oligos for Illumina®?
- What buffers are recommended for shearing DNA in NEB’s enzymatic methyl-seq (EM-seq™) workflows?
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Enzymatic Methyl-seq: The Next Generation of Methylome Analysis
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
