NEBNext^® Enzymatic Methyl-seq Conversion Module

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
NEB # Stored at (°C) E7125S -20 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 Unmethylated Lambda E7123AVIAL -20 1 x 0.024 ml 2 ng/µl 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 E8013AVIAL -20 3 x 100 µl Not Applicable Oxidation Supplement E7128AVIAL -20 1 x 0.024 ml Not Applicable Oxidation Enhancer E7129AVIAL -20 1 x 0.024 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 APOBEC Reaction Buffer E7134AVIAL -20 1 x 0.24 ml Not Applicable BSA E7135AVIAL -20 1 x 0.024 ml Not Applicable 100mM DTT E7139AAVIAL -20 1 x 0.5 ml 100 mM
NEB # Stored at (°C) E7125L -20 Component Name Component # Stored at (°C) Amount Concentration Control DNA CpG methylated pUC19 E7122AAVIAL -20 1 x 0.096 ml 0.1 ng/µl Control DNA Unmethylated Lambda E7123AAVIAL -20 1 x 0.096 ml 2 ng/µl 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 E8013AAVIAL -20 3 x 400 µl Not Applicable Oxidation Supplement E7128AAVIAL -20 1 x 0.096 ml Not Applicable Oxidation Enhancer E7129AAVIAL -20 1 x 0.096 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 APOBEC Reaction Buffer E7134AAVIAL -20 1 x 0.96 ml Not Applicable BSA E7135AAVIAL -20 1 x 0.096 ml Not Applicable 100mM DTT E7139AAVIAL -20 1 x 0.5 ml 100 mM

1. It is possible for FE(II) Solution to have a yellow color. Testing has shown no difference in performance based on color.

1. Where can I find protocols for using the NEBNext® Enzymatic Methyl-seq Kit? (NEB #E7120, E7125, E7140)

1. What types of sample can be processed using the NEBNext® Enzymatic Methyl-seq Kit and the Enzymatic Methyl-seq Conversion Module?
2. What are the recommended inputs for the NEBNext® Enzymatic Methyl-seq Kit and the Enzymatic Methyl-seq Conversion Module?
3. What is the difference between the NEBNext Enzymatic Methyl-seq Kit and the Enzymatic Methyl-seq Conversion Module?
4. What buffers are recommended for shearing DNA in NEB’s enzymatic methyl-seq (EM-seq™) workflows?
5. Can the “active” TET2 Buffer be stored longer than 4 months?
6. Can the freshly diluted Fe(II) Solution be stored long term?
7. Why, at some stages of the EM-seq protocol, do the NEBNext Sample Purification Beads behave differently when cleaning up the sample?
8. Can enzymatically fragmented DNA be used as input material for EM-seq™?
9. Can I use NaOH (sodium hydroxide) instead of formamide to denature my DNA prior to the deamination reaction?
10. Is it normal if the Fe(II) solution from the EM-seq product is yellow or a color change is observed?
11. Can I use NaOH (sodium hydroxide) instead of formamide to denature my DNA before the deamination reaction?
12. Is it normal if the Fe(II) solution from the EM-seq™ product is yellow or a color change is observed?

• 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

• Enzymatic Methyl-seq: The Next Generation of Methylome Analysis

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]

• E7125S_L_G_v1
• E7125S_L_v3

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_v1_10038207
• E7125L_v1_10038235
• E7125S_v1_10045802
• E7125L_v1_10046995
• E7125S_v1_10047944
• E7125S_v1_10038221
• E7125G_v1_10050154
• E7125L_v1_10050475
• E7125S_v1_10050477
• E7125G_v1_10051211
• E7125L_v1_10050867
• E7125S_v1_10050809
• E7125L_v1_10053882
• E7125S_v1_10054046
• E7125G_v1_10058602
• E7125L_v1_10058686
• E7125S_v1_10058692
• E7125S_v1_10062773
• E7125S_v1_10053388
• E7125L_v1_10053399
• E7125G_v2_10069637
• E7125L_v2_10069602
• E7125S_v2_10069600
• E7125L_v2_10070427
• E7125S_v2_10072615
• E7125G_v2_10072933
• E7125L_v2_10072522
• E7125S_v2_10072993
• E7125L_v2_10075500
• E7125L_v2_10080332
• E7125S_v2_10083191
• E7125L_v2_10067511
• E7125G_v2_10083144
• E7125G_v2_10089162
• E7125L_v2_10089166
• E7125S_v2_10089164
• E7125L_v2_10090375
• E7125S_v2_10091590
• E7125L_v2_10083283
• E7125S_v2_10094280
• E7125S_v2_10098379
• E7125L_v2_10101421
• E7125S_v2_10102962
• E7125L_v2_10104052
• E7125G_v2_10104447
• E7125S_v2_10104458
• E7125L_v2_10105642
• E7125S_v2_10106324
• E7125G_v2_10106968
• E7125L_v2_10108663
• E7125G_v2_10110739
• E7125S_v2_10096123
• E7125L_v2_10096136
• E7125L_v2_10117047
• E7125S_v2_10117049
• E7125G_v2_10096098
• E7125L_v2_10125513
• E7125S_v2_10128228
• E7125S_v2_10135546
• E7125L_v2_10139582
• E7125L_v2_10144228
• E7125G_v2_10144680
• E7125S_v2_10144677
• E7125G_v2_10146237
• E7125L_v2_10147664
• E7125S_v2_10148423
• E7125L_v2_10149542
• E7125L_v2_10150205
• E7125L_v2_10127093
• E7125S_v2_10152243
• E7125G_v2_10155162
• E7125L_v2_10155603
• E7125S_v2_10155609
• E7125S_v2_10142135
• E7125G_v2_10142093
• E7125L_v2_10142118
• E7125L_v2_10167691
• E7125S_v2_10167689
• E7125L_v2_10169618
• E7125S_v2_10171499
• E7125L_v2_10171963
• E7125S_v2_10176360
• E7125G_v2_10177627
• E7125L_v2_10158720
• E7125G_v2_10180866
• E7125L_v2_10182012
• E7125S_v2_10158730
• E7125S_v2_10182342
• E7125S_v2_10172392
• E7125G_v2_10158695
• E7125L_v2_10186425
• E7125L_v2_10172376
• E7125G_v2_10172357
• E7125L_v2_10198777
• E7125S_v2_10198782
• E7125L_v2_10201095
• E7125S_v2_10204422
• E7125L_v2_10188607
• E7125L_v2_10201651
• E7125G_v2_10216533
• E7125S_v2_10215759
• E7125S_v2_10201676
• E7125S_v2_10188622
• E7125G_v2_10201621
• E7125S_v2_10212684
• E7125L_v2_10232707
• E7125S_v2_10241783
• E7125L_v2_10232855
• E7125S_v2_10232185
• E7125G_v2_10232173
• E7125L_v2_10256100
• E7125S_v2_10247394
• E7125G_v2_10247352
• E7125L_v2_10266571
• E7125L_v2_10277393
• E7125S_v2_10278173
• E7125G_v2_10212657
• E7125L_v2_10282976
• E7125G_v2_10258921
• E7125S_v2_10258939
• E7125L_v2_10293637
• E7125L_v2_10301741
• E7125S_v2_10301739
• E7125L_v2_10304683
• E7125S_v2_10304686
• E7125L_v2_10326277
• E7125S_v2_10315854
• E7125L_v2_10315845
• E7125L_v2_10331863
• E7125S_v3_10331869

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 Unmethylated Lambda
• Elution Buffer
• TET2 Reaction Buffer
• Oxidation Supplement
• Oxidation Enhancer
• TET2
• Fe (II) Solution
• Stop Reagent
• APOBEC
• APOBEC Reaction Buffer
• BSA
• 100mM DTT

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.