T7 Endonuclease 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

Companion Products

• EnGen® sgRNA Synthesis Kit, S. pyogenes

Materials Sold Separately

• NEBuffer 2

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.

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.

1. Determining Genome Targeting Efficiency using T7 Endonuclease I

• DNA Damage and PreCR

• Activities of DNA Repair Enzymes and Structure-specific Endonucleases
• DNA Repair Enzymes on Damaged and Non-standard Bases
• Properties of DNA Repair Enzymes and Structure-specific Endonucleases
• Properties of Exonucleases and Non-specific Endonucleases

• Exo Selector

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?

• The effect of nucleic acid modifications on digestion by DNA exonucleases

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 & Change Notifications

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

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]

• M0302L_v1_10012368
• M0302S_v1_10012369
• M0302L_v1_10019628
• M0302S_v1_10019630
• M0302L_v1_10022444
• M0302S_v1_10022447
• M0302L_v1_10030738
• M0302L_v1_10034855
• M0302S_v1_10034796
• M0302L_v1_10041804
• M0302S_v1_10041806
• M0302L_v1_10046233
• M0302S_v1_10049332
• M0302L_v1_10051844
• M0302S_v1_10050173
• M0302L_v1_10056681
• M0302S_v1_10056682
• M0302L_v1_10061062
• M0302S_v1_10062736
• M0302L_v1_10064189
• M0302S_v1_10067697
• M0302L_v1_10069250
• M0302L_v1_10076497
• M0302L_v1_10080908
• M0302S_v1_10078800
• M0302L_v1_10082598
• M0302S_v1_10083177
• M0302L_v1_10085720
• M0302S_v1_10088014
• M0302L_v1_10088652
• M0302S_v1_10088654
• M0302L_v1_10095158
• M0302L_v1_10098191
• M0302S_v1_10098318
• M0302L_v1_10106771
• M0302S_v1_10106772
• M0302L_v1_10112773
• M0302S_v1_10116318
• M0302L_v1_10119206
• M0302L_v1_10125107
• M0302S_v1_10125003
• M0302L_v1_10128445
• M0302S_v1_10135670
• M0302L_v1_10134297
• M0302L_v1_10143515
• M0302S_v1_10143517
• M0302L_v1_10151472
• M0302S_v1_10154049
• M0302L_v1_10159639
• M0302S_v1_10159640
• M0302L_v1_10168767
• M0302L_v1_10176782
• M0302S_v1_10176583

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• T7 Endonuclease I
• NEBuffer™ 2

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.