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  • BsaI

    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.

    BsaI has a High Fidelity (HF) Version available.
    cloned at neb recombinant incubation temp heat inactivation cpg dcm
    Bsa-I-cutsite_1
    Catalog #SizeConcentrationPriceQtyAdd to Cart
    R0535S1,000 units10,000 units/ml$63.00Add to Cart
    R0535L5,000 units10,000 units/ml$252.00Add to Cart
      
    Categories:
    Restriction Endonucleases: B
    Applications:
    Golden Gate Assembly,
    Restriction Enzyme Digestion

    Description

    BsaI has a High Fidelity version BsaI-HF™ (NEB #R3535).

    High Fidelity (HF ™) Restriction Enzymes have 100% activity in CutSmart ™ Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. HF enzymes also exhibit dramatically reduced star activity. HF enzymes are all Time-Saver qualified and can therefore cut substrate DNA in 5-15 minutes with the flexibility to digest overnight without degradation to DNA. Engineered with performance in mind, HF restriction enzymes are fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design.

    Product Source

    An E. coli strain that carries the cloned BsaJI gene from Bacillus stearothermophilus 6-55 (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 digest 1 µg of pXba DNA 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 NEBuffers2

    NEBuffer 1.1: 75%
    NEBuffer 2.1: 75%
    NEBuffer 3.1: 100%
    CutSmart™ Buffer: 100%

    Diluent Compatibility

    Storage Temperature

    -20°C

    Storage Conditions

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

    Heat Inactivation

    65°C for 20 min

    Methylation Sensitivity

    dam methylation: Not Sensitive
    dcm methylation: Blocked by Overlapping
    CpG Methylation: Blocked by Some Combinations of Overlapping

    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.
    • Endonuclease Activity (Nicking):
      The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
    • 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.

    Notes

    1. Blocked by overlapping dcm methylation. Cleavage of mammalian genomic DNA is blocked by some combinations of overlapping CpG methylation.
    2. The addition of BSA to the restriction digest allows BsaI to be used at 37°C. This is also true for older lots of BsaI as there has been no formulation change in the enzyme. Activity at 50°C is 100%.
    3. More information about: Methylation Sensitivity
    4. Star activity may result from a glycerol concentration of >5%

    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. 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?
    4. Which restriction enzymes are used in Golden Gate Assembly?
    5. Which restriction enzymes are used in GoldenBraid Assembly?
    1. Optimizing Restriction Endonuclease Reactions
    2. Double Digest Protocol with Standard Restriction Enzymes

    Selection Tools

    Usage Guidelines & Tips

    Troubleshooting Guides

    Interactive Tools

    Citations

    • Feng Y, Zhang S, Huang X (2014)A robust TALENs system for highly efficient mammalian genome editing Sci Rep 4, 3632. PubMedID: 24407151, DOI: 10.1038/srep03632
    • Abil Z, Denard CA, Zhao H (2014)Modular assembly of designer PUF proteins for specific post-transcriptional regulation of endogenous RNA J Biol Eng 8(1), 7. PubMedID: 24581042, DOI: 10.1186/1754-1611-8-7
    • Binder A, Lambert J, Morbitzer R, Popp C, Ott T, Lahaye T, Parniske M (2014)A Modular Plasmid Assembly Kit for Multigene Expression, Gene Silencing and Silencing Rescue in Plants PLoS One 9(2), e88218. PubMedID: 24551083, DOI: 10.1371/journal.pone.0088218
    • Alejandro Sarrion-Perdigones, Marta Vazquez-Vilar, Jorge Palací, Bas Castelijns, Javier Forment, Peio Ziarsolo, José Blanca, Antonio Granell, Diego Orzaez (2013)GoldenBraid 2.0: A Comprehensive DNA Assembly Framework for Plant Synthetic Biology Plant Physiology 162(3), 1618-31. PubMedID: 23669743