NEBNext^® Enzymatic Methyl-seq v2 Conversion Module

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
E8020S     -20       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   NEBNext® Carrier DNA E3351AVIAL -20 1 x 0.024 ml Not Applicable   Elution Buffer E7124AVIAL -20 1 x 2.1 ml Not Applicable   TET2 Reaction Buffer E7126AVIAL -20 1 x 0.3 ml Not Applicable   TET2 Reaction Buffer Supplement (x 1) E8013AVIAL -20 3 x 100 µl Not Applicable   UDP-Glucose E3353AVIAL -20 1 x 0.024 ml Not Applicable   DTT E7139AAVIAL -20 1 x 0.5 ml 100 mM   T4-BGT E3354AVIAL -20 1 x 0.024 ml Not Applicable   T4-BGT Diluent E8014AVIAL -20 1 x 0.216 ml Not Applicable   TET2 E7130AVIAL -20 1 x 0.096 ml Not Applicable   Fe (II) Solution E7131AVIAL -20 1 x 0.024 ml Not Applicable   Stop Reagent E7132AVIAL -20 1 x 0.024 ml Not Applicable   APOBEC E7133AVIAL -20 1 x 0.024 ml Not Applicable   Deamination Reaction Buffer E3356AVIAL -20 1 x 0.096 ml Not Applicable   Recombinant Albumin E3357AVIAL -20 1 x 0.024 ml Not Applicable
E8020L     -20       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   NEBNext® Carrier DNA E3351AAVIAL -20 1 x 0.096 ml Not Applicable   Elution Buffer E7124AAVIAL -20 1 x 8.6 ml Not Applicable   TET2 Reaction Buffer E7126AAVIAL -20 1 x 1.2 ml Not Applicable   TET2 Reaction Buffer Supplement (x 1) E8013AAVIAL -20 3 x 400 µl Not Applicable   UDP-Glucose E3353AAVIAL -20 1 x 0.096 ml Not Applicable   DTT E7139AAVIAL -20 1 x 0.5 ml 100 mM   T4-BGT E3354AAVIAL -20 1 x 0.096 ml Not Applicable   T4-BGT Diluent E8014AAVIAL -20 1 x 0.864 ml Not Applicable   TET2 E7130AAVIAL -20 1 x 0.384 ml Not Applicable   Fe (II) Solution E7131AAVIAL -20 1 x 0.096 ml Not Applicable   Stop Reagent E7132AAVIAL -20 1 x 0.096 ml Not Applicable   APOBEC E7133AAVIAL -20 1 x 0.096 ml Not Applicable   Deamination Reaction Buffer E3356AAVIAL -20 1 x 0.384 ml Not Applicable   Recombinant Albumin E3357AAVIAL -20 1 x 0.096 ml Not Applicable

Companion Products

• NEBNext^® Enzymatic Methyl-seq v2 Kit
• NEBNext^® Enzymatic 5hmC-seq Kit
• NEBNext^® Enzymatic 5hmC-seq Conversion Module
• NEBNext^® Magnetic Separation Rack
• NEBNext^® LV Unique Dual Index Primers Set 2A
• NEBNext^® Primers for Epigenetics (Unique Dual Index Set 2B)
• NEBNext^® LV Unique Dual Index Primers Set 1
• NEBNext^® LV Unique Dual Index Primers Set 2
• NEBNext^® Primers for Epigenetics (Unique Dual Index Set 3)
• NEBNext^® LV Unique Dual Index Primers Set 4
• NEBNext^® LV Unique Dual Index Primers Set 5

1. Where can I find protocols for use of the NEBNext^® Enzymatic Methyl-seq v2 Conversion Module? (NEB #E8020)

• NEBNext Selector

1. What is the difference between the NEBNext^® Enzymatic Methyl-seq v2 Kit (NEB #E8015) and the Enzymatic Methyl-seq v2 Conversion Module (NEB #E8020)?
2. What types of samples can be processed using the NEBNext^® Enzymatic Methyl-seq v2 Kit (NEB #E8015) and the Enzymatic Methyl-seq v2 Conversion Module (NEB #E8020)?
3. What are the recommended inputs for the NEBNext^® Enzymatic Methyl-seq v2 Kit (NEB #E8015) and the Enzymatic Methyl-seq v2 Conversion Module (NEB #E8020)?
4. What buffers are recommended for shearing DNA in NEBNext^® Enzymatic Methyl-seq (EM-seq^™) workflows?
5. What is the concentration of the EM-seq^™ Adaptor and the NEBNext^® LV Unique Dual Index Primers?
6. Can the “active” TET2 Buffer be stored longer than 4 months?
7. Can the freshly diluted Fe(II) Solution be stored long term?
8. Can the freshly diluted T4-BGT be stored long term?
9. What conversion levels are typical with the control DNAs supplied in the EM-seq^™ v2 kits?
10. Can I use NaOH (sodium hydroxide) instead of formamide to denature my DNA before the deamination reaction?
11. Is it normal if the Fe(II) solution from the EM-seq^™ product is yellow or a color change is observed?
12. How can pre-shear spike-in controls be added to cfDNA or other pre-fragmented DNA (amplicons, restriction digested DNA, etc.) going into EM-seq^™ workflow?
13. How should controls be diluted for applications involving shallow sequencing?

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.

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]

• E8020S_L_G_v1

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]

• E8020G_v1_10267963
• E8020L_v1_10267961
• E8020S_v1_10249481

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
• DTT
• TET2
• Fe (II) Solution
• Stop Reagent
• APOBEC

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.

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 busdev@neb.com.