For improved performance, NEBNext Enzymatic Methyl-seq v2 Conversion Module (NEB #E8020) is our recommendation.
NEBNext® Enzymatic Methyl-seq (EM-seq™) is a new method for identification of 5-mC and 5-hmC.
While bisulfite sequencing has been the gold standard for methylome analysis, this conversion treatment damages DNA, resulting in fragmentation, loss and bias. The NEBNext Enzymatic Methyl-seq Conversion Module provides an enzyme-based alternative to bisulfite conversion.
For use in Illumina® sequencing workflows, we recommend use of the NEBNext Enzymatic Methyl-seq Kit (NEB #E7120) instead of this module. The full kit includes the EM-seq Adaptor, which has been optimized for high performance in the EM-seq workflow, as well as the high-efficiency Ultra II library prep reagents, and these components are not included in the Conversion Module.
The NEBNext Enzymatic Methyl-seq Conversion Module provides a high-performance enzyme-based alternative to bisulfite conversion for methylome analysis.
In the EM-seq workflow, TET2 oxidizes 5-mC and 5-hmC, providing protection from deamination by APOBEC in the next step. In contrast, unmodified cytosines are deaminated to uracils.
Features:
High efficiency conversion
Minimal DNA damage
Minimized GC bias
5-mC and 5-hmC detection
Complete kit including library prep reagents also available (NEB #E7120)
For specific detection of 5hmC, the NEBNext Enzymatic E5hmC-seq Conversion Module (NEB #E3365) is now also available.
Figure 1: NEBNext Enzymatic Methyl-seq and Sodium Bisulfite Conversion Methods
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Use this tool to find citations related to NEBNext products. Search by product, keyword or instrument name – use the filters to select specific product areas.
Additional Citations
Sun Z, et al. (2019) Non-destructive enzymatic deamination enables single molecule long read sequencing for the determination of 5-methylcytosine and 5-hydroxymethylcytosine at single base resolution. bioRxiv; DOI: 10.1101/2019.12.20.885061
Vaisvila R, et al. (2021) Enzymatic methyl sequencing detects DNA methylation at single-base resolution from picograms of DNA. Genome Res; PubMedID: 34140313, DOI: 10.1101/gr.266551.120
Publications
Vaisvila R, et al. (2021) Enzymatic methyl sequencing detects DNA methylation at single-base resolution from picograms of DNA. Genome Res; PubMedID: 34140313, DOI: 10.1101/gr.266551.120
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Licenses
In the United States and Europe, this product cannot be used in, or incorporated into, any blood-based, sequencing-based screening diagnostic assay (including product or service-based offerings) for the early detection of colorectal, gastric, lung, liver, ovarian, breast, prostate, esophageal, thyroid, uterine, pancreatic, bladder or kidney cancer indications.
This product and/or its manufacture and/or its use either alone or in combination with other product(s) are protected by one or more of the following patents and related pending patents: U.S. Pat. Nos. 9,121,061, 9,896,726, 10,227,646, 10,260,088, 10,619,200, 11,001,876, and 11,124,825; European Pat. Nos. EP2825645 and EP3368688; JP6224689; and CN108699598.
This product is licensed for research and commercial use from Bio-Rad Laboratories, Inc., under U.S. Pat. No. 8,470,573 and corresponding patents in other countries. No rights are granted for use of the product for Digital PCR or real-time PCR applications, with the exception of quantification in Next Generation Sequencing workflows. For more information, please email [email protected].