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    Nb.BsrDI

    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
    NbBsrDI_cutsite
    Isoschizomers | Single Letter Code
    Catalog #SizeConcentrationPriceQtyAdd to Cart
    R0648S1,000 units10,000 units/ml$65.00Add to Cart
    R0648L5,000 units10,000 units/ml$260.00Add to Cart
    Categories:
    Nicking Endonucleases,
    Restriction Endonucleases: N-O

    Description

    Nb.BsrDI is a nicking endonuclease that cleaves only one strand of DNA on a double-stranded DNA substrate.

    Product Source

    An E. coli strain expressing only the large subunit of the BsrDI restriction gene from Bacillus stearothermophilus D70 (Z. Chen).

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    CutSmart Buffer-2010X

    Properties and Usage

    Unit Definition

    One unit is defined as the amount of enzyme required to convert 1 µg of supercoiled pUC19 DNA to open circular form in 1 hour at 65°C in a total reaction volume of 50 µl.

    Reaction Conditions

    1X CutSmart™ Buffer
    Incubate at 65°C

    1X CutSmart™ Buffer:
    50 mM Potassium Acetate
    20 mM Tris-acetate
    10 mM Magnesium Acetate
    100 μg/ml BSA
    pH 7.9 @ 25°C

    Activity in NEBuffers

    NEBuffer 1.1: 25%
    NEBuffer 2.1: 100%
    NEBuffer 3.1: 100%
    CutSmart™ Buffer: 100%

    Diluent Compatibility

    Storage Temperature

    -20°C

    Storage Conditions

    10 mM Tris-HCl
    200 mM NaCl
    1 mM DTT
    0.1 mM EDTA
    50% Glycerol
    200 μg/ml BSA
    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: 20%

    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.
    • 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.

    Notes

    1. Incubation at 37°C results in 20% activity.

    References

    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]

    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]

    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.

    Datacards

    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. My enzyme used to come with another NEBuffer, but CutSmart™ Buffer is now recommended?  Why?
    2. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
    3. How can I access the old NEBuffer Activity Chart?
    4. How can I access the old Double Digest Finder?
    1. Optimizing Restriction Endonuclease Reactions

    Selection Tools

    Usage Guidelines & Tips

    Troubleshooting Guides

    Interactive Tools