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  • Nb.BtsI

    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
    NbBtsI-cutsite
    Catalog #SizeConcentrationPriceQtyAdd to Cart
    R0707S1,000 units10,000 units/ml$65.00Add to Cart
    R0707L5,000 units10,000 units/ml$260.00Add to Cart
      
    Categories:
    Nicking Endonucleases,
    Restriction Endonucleases: N-O
    Applications:
    DNA Nicking

    Description

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

    Product Source

    An E. coli strain that expresses only the large subunit of the BtsI restriction gene from Bacillus thermoglucosidasius (X.Pan).

    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 plasmid ΦX174 RF I DNA to open circular form in 1 hour at 37°C in a total reaction volume of 50 µl.

    Reaction Conditions

    1X CutSmart™ Buffer
    Incubate at 37°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: 75%
    NEBuffer 2.1: 100%
    NEBuffer 3.1: 75%
    CutSmart™ Buffer: 100%

    Diluent Compatibility

    Storage Temperature

    -20°C

    Storage Conditions

    10 mM Tris-HCl
    50 mM NaCl
    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

    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. Nb.BtsI is very efficient. Under most conditions a one to two hour incubation with 1 µl of enzyme and 1 µg of DNA is recommended. 
    2. Nb.BtsI is 100% active at 55°C. 
    3. To run on an electrophoresis gel, add loading dye containing a final concentration of 0.4% SDS.

    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

    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. 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?
    1. Optimizing Restriction Endonuclease Reactions

    Selection Tools

    Usage Guidelines & Tips

    Troubleshooting Guides

    Interactive Tools

    Also has 100% activity in NEBuffer 2
    Add 0.4% SDS before running agarose gel to dissociate DNA-enzyme complex