Restriction Endonucleases

The Leader in the Discovery and Production of Restriction Enzymes

With over 40 years of offering restriction enzymes to the research community, NEB has earned the reputation of being a leader in enzyme technologies. Working continuously to be worthy of that distinction, NEB strives to develop enzymes of the highest purity and unparalleled performance.

NEB scientists continue to improve its portfolio of restriction enzymes, as well as explore their utility in new technologies. As a result, NEB scientists continue to publish scientific papers and to be awarded grants in this area. With the industry’s largest research and development group dedicated to restriction enzymes, we are proud to have been there first: the first to commercialize a recombinant enzyme, the first to introduce a nicking enzyme, and the first to supply a true restriction enzyme master mix. In addition, NEB has an ongoing history of innovation by engineering restriction enzymes with altered specificities and improved performance. Through continued research in these areas, we are committed to driving the innovations that allow us to offer maximum convenience and performance.

For details on NEB’s quality controls for restriction endonucleases, visit our Restriction Enzyme Quality page.

All of NEB's Restriction enzymes have transitioned to a new buffer system. Visit for further details.


  • >215 restriction enzymes are 100% active in a single buffer – CutSmart™ Buffer.
  • >195 restriction enzymes are Time-Saver qualified, meaning you can digest DNA in 5-15 minutes, or digest DNA safely overnight.
  • Choose from >285 restriction enzymes, the largest selection commercially available.


  • Choose a High-Fidelity (HF®) restriction enzyme, which has been engineered for reduced star activity, rapid digestion (5-15 minutes) and 100% activity in CutSmart Buffer. A vial of 6X Purple Loading Dye is included with most restriction enzymes.
  • All of our restriction enzymes undergo stringent quality control testing, ensuring the highest levels of purity and lot-to-lot consistency.

    Physical Purity Enzymes are evaluated by SDS-PAGE and silver-stained gels to ensure the highest levels of purity and the absence of contaminating proteins.
    DNA Contamination Enzymes are screened by qPCR to ensure no contaminating genomic DNA is present. The specification for this assay is less than one E.coli genome per sample.
    Exonuclease Activity Using radioactively labelled DNA substrate and/or state-of-the-art capillary electrophoresis-based assays with fluorescently-labelled substrates, NEB is able to detect very low levels of exonuclease activity.
    Endonuclease Activity To ensure that there are no contaminating enzymes that could cause nickng or non-specific nuclease degradation, reagents are incubated with supercoiled plasmid DNA for 4 hours to demonstrate the absence of endonuclease contamination. 
    Non-Specific DNase Activity Enzymes are incubated overnight with Lambda DNA to confirm that there is no additional non-specific nuclease activity present. 
    Cloning QC (Ligation and Re-cutting) A DNA template is over-digested by the appropriate restriction enzyme and the percentage of DNA fragments ligated and re-cut are determined by agarose gel electrophoresis.
    Cloning QC (Blue-white Screening) A DNA plasmid is over-digested by the appropriate restriction enzyme and the linearized plasmid DNA is ligated and transformed into an E.coli strain with greater than 99% correct transformants, as determined by a blue-white screen. 

Use Enzyme Finder to select restriction enzyme by name, sequence, overhang or type. 


Restriction Endonucleases includes these subcategories:

Restriction Endonucleases

FAQs for Restriction Endonucleases

Protocols for Restriction Endonucleases

    Publications related to Restriction Endonucleases

  1. Shah, S., Sanchez, J., Stewart, A., et al. 2015. Probing the Run-On Oligomer of Activated SgrAI Bound to DNA PLoS One. 10(4), PubMedID: 25880668, DOI: 10.1371/journal.pone.0124783.
  2. Loenen, W.A., Raleigh, E.A. 2014. The other face of restriction: modification-dependent enzymes. Nucleic Acids Res. 42, PubMedID: 23990325, DOI:
  3. Kamps-Hughes, N., Quimby, A., Zhu, Z., Johnson, E.A. 2013. Massively parallel characterization of restriction endonucleases Scopus. 41(11), PubMedID: 23605040, DOI: 10.1093/nar/gkt257
  4. Fu YB, Peterson G. W., Dong Y 2016. Increasing Genome Sampling and Improving SNP Genotyping for Genotyping-by-Sequencing with New Combinations of Restriction Enzymes G3. 6:4, PubMedID: 26818077, DOI:

Legal Information

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at

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.

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    NEB TV Episode 15

    These days, restriction enzymes are being used in many more applications other than cloning. Learn more in this episode of NEB TV.

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    Type II Restriction Enzymes

    Type II restriction enzymes are most commonly used for molecular biology applications, as they recognize stereotypical sequences and produce a predictable cleavage pattern. Learn more about how Type II REs work.

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    Type I Restriction Enzymes

    Type I restriction enzymes are a group of endonucleases that recognize a bipartite sequence, but do not produce a predictable cleavage pattern. Learn more about how Type I REs work.

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    Type III Restriction Enzymes

    Type III restriction enzymes are a group of endonucleases that recognize a non-pallindromic sequence, comprising two inversely oriented sites. Learn more about these poorly understood enzymes.

  5. Restriction Enzyme Interaction with DNA

    The Interaction of Restriction Enzymes and DNA

    Watch as Geoff Wilson, Restriction Enzyme Division Head, describes the interaction of restriction enzymes and substrate DNA using computer models generated from x-ray crystallography data.

  6. Discovering New Restriction Enzymes

    Discovering New Restriction Enzymes

    Watch as Rick Morgan, Research Scientist in the Restriction Enzyme Division, describes his passion for discovering and characterizing restriction enzymes from nature.

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    Restriction Enzymes in Chromatin Conformation Capture

    Chromatin conformation capture (3C) techniques allow study of the spatial organization of eukaryotic chromosomes in a 3D context. Learn more about this and other applications of restriction enzymes.

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    Restriction Enzymes in Droplet Digital PCR

    Droplet digital PCR is a method for accurately quantitating copies of DNA or RNA in a sample. Each PCR reaction is separated into thousands or millions of droplets for analysis. Learn more about droplet digital PCR.

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    Restriction Enzymes in Golden Gate Assembly

    Type IIS restriction enzymes have both recognition and binding sites, but cut downstream of the recognition site, creating 4-base overhangs ideal for reassembly. View a list of TypeIIS enzymes

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    Restriction Enzymes in Isothermal Amplification

    Isothermal amplification generates many copies of a target sequence in a short period of time, at a constant temperature. Learn more about isothermal amplification.

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    Restriction Enzymes in Optical Mapping

    Optical mapping is a method that allows production of restriction maps of whole chromosomes or genomes. Learn more about optical mapping.