Product Class: Other

Tth Endonuclease IV
NEBU cloned at NEB recombinant 65 No

The large size of this product has been discontinued.

Product Introduction

  • Thermostable DNA AP endonuclease
  • Catalyzes the cleavage of DNA phosphodiester backbone at AP sites via hydrolysis, leaving a 1 nucleotide gap with 3'-hydroxyl and 5' deoxyribose phosphate (dRP) termini
  • Also has 3'-diesterase activity which can remove 3' phosphate, 3'-α, β-unsaturated aldehyde, phosphoglycoaldehyde, and other 3' blocking groups
  • Heat inactivation not possible
Catalog # Size Concentration
M0294S 500.0 units 10000 units/ml

Product Information

Description

Product Source

An E. coli strain that carries the cloned Thermus thermophilus endonuclease IV gene.
This product is related to the following categories:

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

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 (1)
  • Alkaline unwinding (2)

Product Notes

  1. Place reaction in Thermocycler or overlay with mineral oil to reduce evaporation. 
  2. Enzyme stability above 80°C is assured by adding ZnCl2 to a final concentration of 25 μM in the reaction.
  3. Not recommended in NEBuffers 1-4 and ThermoPol® II Reaction Buffer. 
  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. Pflaum, M. et al. (1998). FreeRad. Res.. 29, 585-594.
  2. Hartwig, A. et al. (1996). Toxicology Letters. 88, 85-90.

Protocols, Manuals & Usage

Protocols

  1. Protocol for cleavage with Tth Endonuclease IV (NEB #M0294)

Usage & Guidelines

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. How does Tth Endonuclease IV perform in PCR?

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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