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NEBNext® Enzymatic Methyl-seq Conversion Module

Catalog # Concentration Size List Price Quantity Your Price
E7125L 96 reactions $989.00
$890.10
E7125S 24 reactions $271.00
$243.90
Catalog # Size List Price Your Price
E7125L 96 reactions $989.00
$890.10
E7125S 24 reactions $271.00
$243.90
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

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.

For specific detection of 5hmC, the NEBNext Enzymatic E5hmC-seq Conversion Module (NEB #E3365) is now also available.

Download Technical Note

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





Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Control DNA CpG methylated pUC19 E7122AVIAL -20 1 x 0.024 ml 0.1 ng/µl
  Control DNA CpG unmethylated Lambda E7123AVIAL -20 1 x 0.024 ml 2 ng/µl
  Elution Buffer E7124AVIAL -20 1 x 2.1 ml
  TET2 Reaction Buffer E7126AVIAL -20 1 x 0.3 ml
  TET2 Reaction Buffer Supplement E7127AVIAL -20 3 x 100 µl 100 X
  Oxidation Supplement E7128AVIAL -20 1 x 0.024 ml
  Oxidation Enhancer E7129AVIAL -20 1 x 0.024 ml
  TET2 E7130AVIAL -20 1 x 0.096 ml
  Fe (II) Solution E7131AVIAL -20 1 x 0.024 ml
  Stop Reagent E7132AVIAL -20 1 x 0.024 ml
  APOBEC E7133AVIAL -20 1 x 0.024 ml
  APOBEC Reaction Buffer E7134AVIAL -20 1 x 0.24 ml
  BSA E7135AVIAL -20 1 x 0.024 ml
  DTT E7139AAVIAL -20 1 x 0.5 ml 100 mM
  Control DNA CpG methylated pUC19 E7122AAVIAL -20 1 x 0.096 ml 0.1 ng/µl
  Control DNA CpG unmethylated Lambda E7123AAVIAL -20 1 x 0.096 ml 2 ng/µl
  Elution Buffer E7124AAVIAL -20 1 x 8.6 ml
  TET2 Reaction Buffer E7126AAVIAL -20 1 x 1.2 ml
  TET2 Reaction Buffer Supplement E7127AAVIAL -20 3 x 400 µl 100 X
  Oxidation Supplement E7128AAVIAL -20 1 x 0.096 ml
  Oxidation Enhancer E7129AAVIAL -20 1 x 0.096 ml
  TET2 E7130AAVIAL -20 1 x 0.384 ml
  Fe (II) Solution E7131AAVIAL -20 1 x 0.096 ml
  Stop Reagent E7132AAVIAL -20 1 x 0.096 ml
  APOBEC E7133AAVIAL -20 1 x 0.096 ml
  APOBEC Reaction Buffer E7134AAVIAL -20 1 x 0.96 ml
  BSA E7135AAVIAL -20 1 x 0.096 ml
  DTT E7139AAVIAL -20 1 x 0.5 ml 100 mM
An exception occurred during the operation, making the result invalid. Check InnerException for exception details.

Notes
  • It is possible for FE(II) Solution to have a yellow color. Testing has shown no difference in performance based on color.
Citations
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 ResPubMedID: 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
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
Legal And Disclaimer

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

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.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.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.

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