Product Class: Other

T7 Endonuclease I
NEB2 Gray property icon for cloned at NEB recombinant Gray property icon for incubation temperature 37° Gray property icon for heat inactivation no

Authenticase® (NEB #M0689) now available for improved genome editing mutation detection.

Product Introduction

Mechanism of T7 Endonuclease I

  • DNA Endonuclease
  • Catalyzes the cleavage of DNA mismatches and non-β DNA structures including Holliday junctions and cruciform leaving 3'-OH and 5'-phosphate
  • Best at C mismatches and does not recognize all DNA mismatches
  • To a lesser extent cleaves across a nick in dsDNA
  • Key enzyme for use in genome editing mutation detection workflows
  • Also available in convenient kit format: EnGen Mutation Detection Kit (NEB #E3321)
  • Need help finding the right exonuclease for your experiments? Try Exo Selector 
Bioz Badge Exists : True
Catalog # Size Concentration
M0302S 250 units 10000 units/ml
M0302L 1250 units 10000 units/ml

Product Information

Description

Highlights

  • Isolated from a recombinant source
  • Supplied with 10X Reaction Buffer
  • Recognizes non-perfectly matched DNA
T7 Endonuclease I recognizes and cleaves non-perfectly matched DNA, cruciform DNA structures, Holliday structures or junctions, heteroduplex DNA and more slowly, nicked double-stranded DNA. The cleavage site is at the first, second or third phosphodiester bond that is 5´ to the mismatch. The protein is the product of T7 gene 3.

Product Source

An E. coli strain that carries a fusion of maltose binding protein and T7 Endonuclease I (T7 Endo I).
This product is related to the following categories:
Validation of CRISPR-based Gene Editing,
Exonucleases and Non-specific Endonucleases,
DNA Repair Enzymes and Structure-specific Endonucleases,
This product can be used in the following applications:
Whole Genome Amplification & Multiple Displacement Amplification

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

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

Properties & Usage

Materials Required but not Supplied

Unit Definition

One unit is defined as the amount of enzyme required to convert > 90% of 1 μg of supercoiled cruciform pUC(AT) to > 90% linear form in a total reaction volume of 50 μl in 1 hour at 37°C.

Reaction Conditions

1X NEBuffer™ 2
Incubate at 37°C

1X NEBuffer™ 2
50 mM NaCl
10 mM Tris-HCl
10 mM MgCl2
1 mM DTT
(pH 7.9 @ 25°C)

Storage Buffer

20 mM Tris-HCl
200 mM NaCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
0.15% Triton® X-100
pH 7.5 @ 25°C

Heat Inactivation

No

Application Features

  • Resolve four-way junction or branched DNA
  • Detect or cleave heteroduplex and nicked DNA
  • Randomly cleave linear DNA for shot-gun cloning

Product Notes

  1. T7 Endonuclease I is a structure-selective enzyme. It acts on a variety of DNA substrates with different specific activities.
  2. It is important to control the amount of enzyme and the reaction time used for cleavage of a particular substrate.
  3. Temperatures above 42°C cause an increase in nonspecific nuclease activity and should be avoided.
  4. pUC(AT) is derived from pUC19 with a modification of the polylinker between the EcoRI site and the PstI site.

References

  1. Xu, M. -Q. and Evans, T.C. (2001). Methods. 24, 257-277.
  2. Parkinson, M.J. and Lilley. D.M.J. (1997). J. Mol. Biol.. 270, 169-178.
  3. White, M.F. et al. (1997). J. Mol. Biol.. 269, 647-664.
  4. Hadden, J.M. et al. (2001). Nat. Struct. Biol.. 8, 62-67.

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. Is there a way to inactivate T7 Endonuclease I?
  2. Does T7 Endonuclease I recognize single base pair mismatches?
  3. What common additives inhibit T7 Endonuclease I?
  4. What is the molecular weight of T7 Endonuclease I?
  5. What PCR reagents do you recommend for DNA amplification in genome editing (CRIPSR/Cas9,TALEN,ZFN) mismatch detection assays?
  6. Can I use T7 Endonuclease I genome editing (CRIPSR/Cas9,TALEN,ZFN) mismatch detection assays using unpurified PCR products?
  7. Can I use T7 Endonuclease I for genome editing applications?
  8. What is the molecular weight of T7 Endonuclease I?
  9. What PCR reagents do you recommend for DNA amplification in genome editing (CRIPSR/Cas9,TALEN,ZFN) mismatch detection assays?

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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