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Tma Endonuclease III

Catalog # Concentration Size List Price Quantity Your Price
M0291S 10000 units/ml 500 units $123.00
$110.70
Catalog # Size List Price Your Price
M0291S 500 units $123.00
$110.70
Catalog #
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

  • 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
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Product Source
Cloned from Thermotoga maritima and overexpressed in E. coli.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Tma Endonuclease III M0291SVIAL -20 1 x 0.05 ml 10,000 units/ml
  ThermoPol® Reaction Buffer B9004SVIAL -20 1 x 1.5 ml 10 X
Application Features
  • Alkaline elution (3)
  • Alkaline unwinding (4)

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.

Materials Sold Separately
Notes
  • Place reaction in Thermocycler or overlay with mineral oil to reduce evaporation. 
  • Tma Endo III has 75% activity in ThermoPol® II (Mg-free) Reaction Buffer (NEB #B9005).
  • 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.
  • 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
  • Nelson, K.E. et al. (1999). Nature. 399,
  • Marks, K. unpublished observation.
  • Pflaum, M. et al. (1998). FreeRad. Res. 29,
  • Hartwig, A. et al. (1996). Toxicol. Lett. 88,
Quality Control Assay
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 Data Sheets
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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.

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