T7 Endonuclease I

Catalog #	Concentration	Size	List Price	Quantity	Your Price
M0302L	10000 units/ml	1250 units	$447.00		$402.30
M0302S	10000 units/ml	250 units	$112.00		$100.80

Catalog #	Size	List Price	Your Price
M0302L	1250 units	$447.00	$402.30
M0302S	250 units	$112.00	$100.80

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.

Highlights

Isolated from a recombinant source
Supplied with 10X Reaction Buffer
Recognizes non-perfectly matched DNA

Product Source

An E. coli strain that carries a fusion of maltose binding protein and T7 Endonuclease I (T7 Endo I).

Reagents Supplied

The following reagents are supplied with this product:

NEB #

Component Name

Component #

Stored at (°C)

Amount

Concentration

M0302S -20

	T7 Endonuclease I	M0302SVIAL	-20	1 x 0.025 ml	10,000 units/ml
	NEBuffer™ 2	B7002SVIAL	-20	1 x 1.25 ml	10 X

M0302L -20

	T7 Endonuclease I	M0302LVIAL	-20	1 x 0.125 ml	10,000 units/ml
	NEBuffer™ 2	B7002SVIAL	-20	1 x 1.25 ml	10 X

Application Features

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

Properties & Usage

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% TritonÂ® X-100
pH 7.5 @ 25°C

Heat Inactivation

Companion Products

EnGen® sgRNA Synthesis Kit, S. pyogenes

Materials Sold Separately

NEBuffer 2

Notes

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

References

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

Protocols

Determining Genome Targeting Efficiency using T7 Endonuclease I

Usage Guidelines

DNA Damage and PreCR

Selection Charts

Web Tools

Exo Selector

FAQs

Citations

See more details on Bioz

Additional Citations

Zheng Hu, Wencheng Ding, Da Zhu, Lan Yu, Xiaohui Jiang, Xiaoli Wang, Changlin Zhang, Liming Wang, Teng Ji, Dan Liu, Dan He, Xi Xia, Tao Zhu, Juncheng Wei, Peng Wu, Changyu Wang, Ling Xi, Qinglei Gao, Gang Chen, Rong Liu, Kezhen Li, Shuang Li, Shixuan Wang, Jianfeng Zhou, Ding Ma, Hui Wang (2015) TALEN-mediated targeting of HPV oncogenes ameliorates HPV-related cervical malignancy. J Clin Invest 125, 425-36.PubMedID: 25500889, DOI: 10.1172/JCI78206
A-Bk Dad, S Ramakrishna, M Song, H Kim (2014) Enhanced gene disruption by programmable nucleases delivered by a minicircle vector. Gene Ther ,PubMedID: 25142139, DOI: 10.1038/gt.2014.76

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]

M0302S_L_v1

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

Legal And Disclaimer

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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