NEBNext® Enzymatic Methyl-seq Conversion Module
Product informationCode | Name | Size | Quantity | Price | |
---|---|---|---|---|---|
E7125S |
NEBNext Enzymatic Methyl-seq Conversion Module |
24 Assays | - | Unavailable in your region | |
E7125L |
NEBNext Enzymatic Methyl-seq Conversion Module |
96 Assays | - | Unavailable in your region |
NEBNext® Enzymatic Methyl-seq Conversion Module
Product Introduction
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.
- High efficiency conversion
- Minimal DNA damage
- Minimized GC bias
- 5-mC and 5-hmC detection
- Complete kit including library prep reagents also available
For specific detection of 5hmC, the NEBNext Enzymatic E5hmC-seq Conversion Module (NEB #E3365) is now also available.
Catalog # | Size | Concentration |
---|---|---|
E7125S | 24.0 reactions | |
E7125L | 96.0 reactions |
- Product Information
- Protocols, Manuals & Usage
- FAQs & Troubleshooting
- Citations & Technical Literature
- Quality, Safety & Legal
- Other Products You May Be Interested In
Product Information
Description
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.
- This product is related to the following categories:
- Automation for NEBNext® NGS Library Prep Products,
- Epigenetic Analysis Products,
- Epigenetics Products,
- This product can be used in the following applications:
- DNA Methylation Analysis,
- Bisulfite Conversion,
- Epigenetics, Methylated DNA Analysis Products
Reagents Supplied
Reagents Supplied
The following reagents are supplied with this product:
Properties & Usage
Materials Required but not Supplied
- PCR strip tubes
- Formamide (Sigma #F9037-100 ml) or 0.1 N NaOH
- 80% Ethanol
- 0.1X TE, pH 8.0
- Nuclease-free Water
- Clean-up Beads: NEBNext Sample Purification Beads (supplied with Enzymatic Methyl-seq Kit, NEB #E7120), SPRIselect® Reagent Kit (Beckman Coulter, Inc. #B23317), AMPure® XP Beads (Beckman Coulter, Inc. #A63881) or preferred bead manufacturer.
- Magnetic rack/stand, such as NEBNext Magnetic Separation Rack (NEB #S1515)
- PCR machine
- Bioanalyzer®, TapeStation® and associated consumables or other fragment analyzer
Product Notes
- It is possible for FE(II) Solution to have a yellow color. Testing has shown no difference in performance based on color.
Protocols, Manuals & Usage
Protocols
Manuals
FAQs & Troubleshooting
FAQs
- What types of sample can be processed using the NEBNext® Enzymatic Methyl-seq Kit and the Enzymatic Methyl-seq Conversion Module?
- What are the recommended inputs for the NEBNext® Enzymatic Methyl-seq Kit and the Enzymatic Methyl-seq Conversion Module?
- What is the difference between the NEBNext Enzymatic Methyl-seq Kit and the Enzymatic Methyl-seq Conversion Module?
- What buffers are recommended for shearing DNA in NEB’s enzymatic methyl-seq (EM-seq™) workflows?
- Can the “active” TET2 Buffer be stored longer than 4 months?
- Can the freshly diluted Fe(II) Solution be stored long term?
- Why, at some stages of the EM-seq protocol, do the NEBNext Sample Purification Beads behave differently when cleaning up the sample?
- Can enzymatically fragmented DNA be used as input material for EM-seq™?
- Can I use NaOH (sodium hydroxide) instead of formamide to denature my DNA prior to the deamination reaction?
- Is it normal if the FE(II) solution from the EM-seq product is yellow or a color change is observed?
Citations & Technical Literature
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 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
Feature Articles
Quality, Safety & Legal
Quality Assurance Statement
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]- E7125G_v0_10043542
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Safety DataSheets
The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.Control DNA CpG methylated pUC19
Control DNA CpG unmethylated Lambda
Elution Buffer
TET2 Reaction Buffer
TET2 Reaction Buffer Supplement
Oxidation Supplement
Oxidation Enhancer
TET2
Fe (II) Solution
Stop Reagent
APOBEC
APOBEC Reaction Buffer
BSA
DTT
Legal and Disclaimers
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.
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.The supporting documents available for this product can be downloaded below.