Product Class: Other

T5 Exonuclease
NEB4 cloned at NEB recombinant 37 No

This product replaces T5 Exonuclease (NEB #M0363).

GMP-grade reagent also available (Learn more).


Catalog #M0663

Product Introduction

  • Double-stranded DNA specific exonuclease and single-stranded DNA endonuclease
  • Initiates at the 5' termini of linear or nicked double-stranded DNA
  • Cleaves linear or nicked double-stranded DNA in the 5' to 3' direction
  • Need help finding the right exonuclease for your experiments? Try Exo Selector 

T5 Exonuclease is ideal for:

  • Removal of incomplete ligation products from ligated circular double-stranded DNA
  • Degradation of denatured DNA from alkaline-based plasmid purification methods for improved DNA cloning
  • Degradation of contaminating linear and nicked DNA in plasmid samples

Product Information

Description

T5 Exonuclease degrades DNA in the 5´ to 3´ direction (1). T5 Exonuclease is able to initiate nucleotide removal from the 5′ termini or at gaps and nicks of linear or circular dsDNA (1). However, the enzyme does not degrade supercoiled dsDNA (2). T5 Exonuclease also has ssDNA endonuclease activity.

Product Source

The T5 phage D15 gene is expressed as His-tag fusion in E. coli.
This product is related to the following categories:
Exonucleases and Non-specific Endonucleases
This product can be used in the following applications:
Gibson Assembly®

Properties & Usage

Unit Definition

1 unit of T5 Exonuclease is defined as the amount of enzyme required to cause the change of  0.00032 A260 nm/min at 37° C in rCutSmart Buffer.

Reaction Conditions

1X NEBuffer™ 4
Incubate at 37°C

1X NEBuffer™ 4
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
1 mM DTT
(pH 7.9 @ 25°C)

Storage Buffer

50 mM Tris-HCl
100 mM NaCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
0.1% Triton® X-100
pH 7.5 @ 25°C

Heat Inactivation

No

Unit Assay Conditions

1X CutSmart Buffer; 40 µg/ml Salmon Sperm DNA

Application Features

  • Degradation of linear ssDNA, dsDNA or nicked plasmid DNA while maintaining supercoiled plasmid DNA.
  • Remove denatured DNA isolated during alkaline lysis-based plasmid purification procedure (5).
  • Enhance the transfection efficiency of miniprep from plasmid cDNA libraries (6).

References

  1. Sayers, J.R. et al. (1991). Nucleic Acids Res. 19, 4127-4132.
  2. Sayers, J.R. et al. (1990). J. of Biol. Chem. 265, 18311-18317.
  3. Kaliman, A.V. et al. (1986). FEBS. 195, 61-64.
  4. Frenkel, G.D. et al. (1971). J. of Biol. Chem. 246, 4839-4847.
  5. Sayers, J.R. et al. (1996). Analytical Biochem. 241, 186-189.
  6. Kiss-Toth, E. et al. (2001). Analytical Biochem. 288, 230-232.

Protocols, Manuals & Usage

Protocols

  1. Protocol for T5 Exonuclease (NEB #M0663)

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

Research Posters

FAQs & Troubleshooting

FAQs

  1. How Does T5 Exonuclease differ from Lambda Exonuclease (NEB# M0262)?
  2. Will T5 exonuclease cleave supercoiled dsDNA?
  3. How does T5 Exonuclease differ from Exonuclease III (NEB# M0206)?
  4. What is the activity of T5 Exonuclease in the NEBuffers?
  5. Can T5 Exonuclease be heat inactivated?
  6. Can DNA be blunted using T5 Exonuclease?
  7. Will T5 Exonuclease work in rCutSmart buffer?
  8. What is the difference between T5 Exonuclease (NEB #M0363) and T5 Exonuclease (NEB #M0663)?

Tech Tips

Has 5’ to 3’ dsDNA exonuclease activity and ssDNA endonuclease activity.

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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