Product Class: Other

Thymine-DNA Glycosylase (TDG)
Gray property icon for NEBU Gray property icon for cloned at NEB recombinant Gray property icon for incubation temperature 37° Gray property icon for heat inactivation 65°

Product Introduction

Image showing mechanism of Thymine-DNA Glycosylase


  • Mutant form of human thymine-DNA glycosylase.
  • Monofunctional DNA glycosylase that catalyzes the hydrolysis of the N-glycosidic bond to release the damaged base.
  • Major substrates include dU:dG or dT:dG mispairs(1).
  • Minor substrates include 5-formylcytosine and 5-carboxylcytosine (oxidation products of 5mC)(2), 5-hydroxymethyluracil(3), 3, N4- ethenocytosine(4), 5-fluorouracil(5), and other modified derivatives(6).
  • Slight preference for mispairs within a CpG context.
Bioz Badge Exists : True
Catalog # Size Concentration
M0766S 100 units 2000 units/ml
M0766L 500 units 2000 units/ml

Product Information

Description

Thymine-DNA glycosylase (TDG) catalyzes the hydrolysis of the N-glycosidic bond of thymine or uracil mispaired opposite guanine that arise from spontaneous deamination events of 5-methylcytosine (5mC) and cytosine, respectively. The excision of the mispaired thymine or uracil base leaves an abasic (AP) site that can then be chemically or enzymatically cleaved. 

Product Source

An E. coli strain that carries a cloned truncated mutant form of the human TDG gene.
This product is related to the following categories:
DNA Repair Enzymes and Structure-specific Endonucleases,

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • NEB # Stored at (°C)
    M0766S -20
  • NEB # Stored at (°C)
    M0766L -20

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to create an AP site from 1 pmol of a 60-mer oligonucleotide duplex containing a single T:G mispair within a CpG context* in a total reaction of 20 µl in 1 hour at 37°C.

*A T:G mispair is prepared by having the T located at the 24th position of a 5′ FAM-labelled 60 mer ssDNA and a
G at the 25th position of an unlabeled complement, annealed to form a duplex substrate.

Reaction Conditions

1X TDG Reaction Buffer
Incubate at 37°C

1X TDG Reaction Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
0.25 mM TCEP
10 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Storage Buffer

10 mM Tris-HCl
500 mM NaCl
0.1 mM EDTA
1 mM DTT
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

65°C for 10 minutes

References

  1. Waters, T.R. and Swann, P.F. (1998). 273, 20007-14. J Biol Chem. PubMedID: 9685338
  2. He Y.F. et al. (2011). 333, 1303-07. Science. PubMedID: 21817016
  3. Baker D. et al. (2002). 15, 33-9. Chem Res Toxicol. PubMedID: 11800595
  4. Saparbaev, M. and Laval, J. (1998). 95, 8508-13. PNAS. PubMedID: 9671708
  5. Hardeland, U. et al. (2000). 275, 33449-33456. J Biol Chem. PubMedID: 10938281
  6. Bennett M.T. et al. (2006). 128(38), 12510-9 . J Am Chem Soc. PubMedID: 16984202

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. Is Thymine-DNA Glycosylase (TDG) (NEB #M0766) a tagged protein?
  2. What is the activity of Thymine-DNA Glycosylase (TDG) (NEB #M0766) in different NEBuffers?
  3. Is Thymine-DNA Glycosylase (TDG) (NEB #M0766) inhibited by salt?
  4. I do not want to use the provided Thymine-DNA Glycosylase (TDG) reaction buffer for my experiment. Do you have any suggestions on how I can optimize my buffer conditions?
  5. Can Thymine-DNA Glycosylase (TDG) (NEB #M0766) be heat-inactivated?
  6. What bases will Thymine-DNA Glycosylase (TDG) (NEB #M0766) recognize and remove?
  7. What substrate is used to test Thymine-DNA Glycosylase (TDG) (NEB #M0766)?

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]

Safety DataSheets

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

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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.