Product Class: Other

Tma Endonuclease III
Gray property icon for NEBU Gray property icon for cloned at NEB recombinant 65 Gray property icon for heat inactivation no

  • Catalog # M0291 was discontinued on December 15, 2024

Product Introduction

  • Thermostable bifunctional DNA glycosylase with DNA N-glycosylase and AP lyase activities
  • The N-glycosylase activity releases damaged pyrimidines, including thymine glycol and 5,6-dihydroxythymine, generating an AP site. The AP lyase activity cleaves an AP site via β-elimination, creating a 1-nucleotide gap with 3'-α, β-unsaturated aldehyde and 5'-phosphate termini.
  • Heat inactivation not possible
Bioz Badge Exists : True

Product Information

Description

 

Product Source

Cloned from Thermotoga maritima and overexpressed in E. coli.
This product is related to the following categories:
Discontinued (<3 years),

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to cleave 1 pmol of a 60-mer oligonucleotide duplex containing a single AP site* in a total reaction volume of 10 µl in 1 hour at 65°C. 

*An AP site is created by treating 10 pmol of a 60-mer oligonucleotide duplex containing a single uracil residue with 1 unit of Uracil-DNA Glycosylase (UDG) for 2 minutes at 37°C. .

Reaction Conditions

1X ThermoPol® Reaction Buffer
Incubate at 65°C

1X ThermoPol® Reaction Buffer
20 mM Tris-HCl
10 mM (NH4)2SO4
10 mM KCl
2 mM MgSO4
0.1% Triton® X-100
(pH 8.8 @ 25°C)

Usage Concentration

10,000 units/ml

Storage Buffer

10 mM Tris-HCl
100 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
0.1% Triton® X-100
pH 7.4 @ 25°C

Heat Inactivation

No

Unit Assay Conditions

1X ThermoPol Reaction Buffer containing 5 pmol of fluorescently labeled oligonucleotide duplex in a total reaction volume of 10 µl.

Application Features

  • Alkaline elution (3)
  • Alkaline unwinding (4)

Product Notes

  1. Place reaction in Thermocycler or overlay with mineral oil to reduce evaporation. 
  2. Tma Endo III has 75% activity in ThermoPol® II (Mg-free) Reaction Buffer (NEB #B9005).
  3. Tma Endo III has a 100% activity in NEBuffer 1 (NEB# B7001), 100% activity in NEBuffer 4 (NEB #B7004), and 50% activity in NEBuffer 2 (NEB #B7002). The use of Tma Endo III in NEBuffer 3 (NEB #B7003) is not recommended.
  4. Diluent Compatibility:   Diluent D:
    100 mM KCl, 10 mM Tris-HCl (pH 7.4 @ 25°C), 0.1 mM EDTA, 1 mM DTT, 0.1% Triton X-100 and 50% glycerol.

References

  1. Nelson, K.E. et al. (1999). Nature. 399,
  2. Marks, K. unpublished observation.
  3. Pflaum, M. et al. (1998). FreeRad. Res. 29,
  4. Hartwig, A. et al. (1996). Toxicol. Lett. 88,

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. What is the activity of Tma Endonuclease III in the NEBuffers 1-4?
  2. What is the activity of Tma Endonuclease III in ThermoPol buffer?
  3. What is the molecular weight of Tma Endonuclease III?
  4. Is Tma Endonuclease III a tagged protein?
  5. What substrate is used to test the Tma Endonuclease III activity?
  6. What types of DNA damage does Tma Endonuclease III act upon?

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.

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.

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