TET2

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
M1524S     -20       TET2 M1524SVIAL -20 1 x 0.05 ml 8,000 units/ml   TET2 Reaction Buffer B5501SVIAL -20 1 x 1.25 ml 5 X   Dithiothreitol (DTT) B1222AVIAL -20 1 x 0.5 ml 100 mM   Fe(II) Solution B5503SVIAL -20 1 x 0.25 ml 500 mM   TET2 Reaction Buffer Supplement B5502SVIAL -20 3 x 100 µl Not Applicable
M1524L     -20       TET2 M1524LVIAL -20 1 x 0.25 ml 8,000 units/ml   TET2 Reaction Buffer B5501SVIAL -20 2 x 1.25 ml 5 X   Dithiothreitol (DTT) B1222AVIAL -20 2 x 0.5 ml 100 mM   Fe(II) Solution B5503SVIAL -20 1 x 0.25 ml 500 mM   TET2 Reaction Buffer Supplement B5502LVIAL -20 3 x 400 µl Not Applicable

Features

• Specifically oxidizes 5mC, to 5hmC, 5fC, and 5caC in a stepwise manner.

1. For enzymes that cannot be heat-inactivated, we recommend using a column for cleanup (such as the Monarch^® PCR & DNA Cleanup Kit, NEB #T1130) or treatment with Thermolabile Proteinase K (NEB #P8111).
2. TET2 is active across a wide temperature range (4-80°C). Optimal temperature is 37°C.
3. Under nonstandard conditions, TET2 activity can become sensitive to sequence context.
4. The methylation status on the opposing strand does not impact TET2 activity.

1. Crawford D, et al. (2016). Tet2 Catalyzes Stepwise 5-methylcytosine Oxidation by an Iterative and de novo Mechanism. 138(3), 730-733. DOI: 10.1021/jacs.5b10554 J. Am. Chem. Soc.
2. Vaisvila R, et al. (2021). Enzymatic methyl sequencing detects DNA methylation at single-base resolution from picograms of DNA. 31(7), 1280-1289. DOI: 10.1101/gr.266551.120 Genome Research.
3. Ravichandran M, et al. (2022). Pronounced sequence specificity of the TET enzyme catalytic domain guides its cellular function. 8(36), DOI: 10.1126/sciadv.abm2427 Science Advances.
4. DeNizio J, et al. (2022). TET-TDG active DNA demethylation at CpG and non-CpG site. 433(8), 166877. DOI: 10.1016/j.jmb.2021.166877 J Mol Bio.
5. WU H, et al. (2014). Reversing DNA Methylation: Mechanisms, Genomics, and Biological Function. 156(1-2), 45-68. DOI: 10.1016/j.cell.2013.12.019 Cell.

1. Oxidation of Methylated Cytosine Using TET2 (NEB #M1524)

1. Can TET2 (NEB #M1524) be heat inactivated?
2. Can the freshly diluted Fe(II) Solution be stored long term?
3. Can the reconstituted 1X TET2 Reaction Buffer Supplement (supplied with NEB #M1524) be stored longer than 4 months?
4. Does TET2 (NEB #M1524) exhibit any sequence preference?
5. How long can the lyophilized TET2 Reaction Buffer Supplement (supplied with NEB #M1524) be stored?
6. Is it normal if the Fe(II) solution is yellow or a color change is observed?
7. Is TET2 (NEB #M1524) active on asymmetrically methylated substrates?
8. Is TET2 (NEB #M1524) active on methylated RNA?
9. Is TET2 (NEB #M1524) active on methylated single stranded DNA?
10. Is TET2 (NEB #M1524) inhibited by EDTA?
11. Is TET2 (NEB #M1524) inhibited by salt?
12. What is the activity of TET2 (NEB #M1524) in other buffers?
13. What is the reaction temperature range for TET2 (NEB #M1524)?

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]

• M1524S_L_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]

• M1524S_v1_10309641
• M1524L_v1_10309639

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• TET2
• Dithiothreitol (DTT)

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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

Notice to Purchaser: No rights are conveyed for the use of this product in NEB’s EM-seq technology, which is covered by patents on compositions and/or methods including US Patent Nos.10,227,646, 10,260,088, 10,619,200, 11,001,876, and 11,124,825; EP3368688; JP6224689; and CN108699598, and corresponding pending patent applications. Use of this product for NEB’s EM-seq technology requires a license from NEB. For more information, please email busdev@neb.com.