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  • Nt.BstNBI

    This enzyme has transitioned to an improved new buffer system. Visit NEBCutSmart.com for further details.
    The new and current Double Digest Finder and current Activity/Performance Chart for the CutSmart buffer system are available. The previous version of the Double Digest Finder, as well as the previous Version of Activity/Performance Chart that use the former buffer system, are still available for your convenience.
    cloned at neb recombinant incubation temp heat inactivation
    Catalog #SizeConcentrationPriceQtyAdd to Cart
    R0607S1,000 units10,000 units/ml$65.00Add to Cart
    R0607L5,000 units10,000 units/ml$260.00Add to Cart
    Nicking Endonucleases,
    Restriction Endonucleases: N-O
    DNA Nicking


    Nt.BstNBI is a site specific endonuclease that cleaves only one strand of DNA on a double-stranded DNA substrate. The nicking endonuclease catalyzes a single strand break 4 bases beyond the 3´ side of the recognition sequence.


    • N.BstNB I catalyzes a single-stranded nick 3' of its recognition sequence.

    Product Source

    An E. coli strain that carries the cloned Nt.BstNBI gene from Bacillus stereothermophilus 33M (Z. Chen).

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    NEBuffer 3.1-2010X

    Properties and Usage

    Unit Definition

    One unit is defined as the amount of enzyme required to digest 1 µg T7 DNA in 1 hour at 55°C in a total reaction volume of 50 µl.

    The phage T7 DNA is a particularly convenient substrate for assaying the activity because of the proximity of GAGTC sequences. Nt.BstNBI cleavage of phage T7 DNA yields 5 bands; 13 kb, 9 kb, 7 kb, 6 kb and 4 kb.

    Reaction Conditions

    1X NEBuffer 3.1
    Incubate at 55°C

    1X NEBuffer 3.1:
    100 mM NaCl
    50 mM Tris-HCl
    10 mM MgCl2
    100 μg/ml BSA
    pH 7.9 @ 25°C

    Activity in NEBuffers

    NEBuffer 1.1: 0%
    NEBuffer 2.1: 10%
    NEBuffer 3.1: 100%
    CutSmart™ Buffer: 10%

    Diluent Compatibility

    Storage Conditions

    10 mM Tris-HCl
    50 mM KCl
    1 mM DTT
    0.1 mM EDTA
    200 μg/ml BSA
    50% Glycerol
    pH 7.4 @ 25°C

    Heat Inactivation

    80°C for 20 min

    Methylation Sensitivity

    dam methylation: Not Sensitive
    dcm methylation: Not Sensitive
    CpG Methylation: Not Sensitive

    Activity at Temperature

    @37°C: 10%

    Quality Control

    Quality Control Assays

    The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
    • Exonuclease Activity (Radioactivity Release):
      The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
    • Ligation and Recutting (Terminal Integrity):
      After an over-digestion of DNA with a restriction endonuclease the percentage of the DNA fragments ligated with T4 DNA ligase and the percentage that can be recut are determined by agarose gel electrophoresis.
    • Non-Specific DNase Activity (16 hour):
      The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.


    1. 10 units of enzyme are required to convert 1 µg of supercoiled pBR322 or pUC19 DNA to open circular form in 1 hour at 55°C.


    1. Song, Q. et al. (2010). Anal. Chem. [Epub ahead of print].
    2. Zhang, P. et al. (2010). Protein Expr. Purif. 69, 226-234. [Epub 2009 Sep 9].

    Supporting Documents

    Certificate of Analysis

    The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]


    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]

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.


    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.
    1. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
    2. How can I access the old NEBuffer Activity Chart?
    3. How can I access the old Double Digest Finder?

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