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BsaI-HFv2 (1000U)

71,00 €
SKU R3733S

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Attention Golden Gate Assembly users: BsaI-HFv2 (NEB #R3733) has been optimized for Golden Gate Assembly*.  BsaI-HFv2 also works well for any protocol requiring DNA cutting by BsaI. This is the recommended enzyme for any purpose requiring digestion at the recognition sequence ...5′-GGTCTC(N1)/(N5)-3′...
*The requirements for digestion of DNA during Golden Gate Assembly are more demanding than what is encountered in traditional DNA cloning. For Golden Gate, the enzyme must function well in a buffer that might not be the same buffer as that normally provided for the restriction enzyme, and maintain activity for an extended time at elevated temperatures in a dynamic cutting/re-ligating reaction with competition for substrate binding between the endonuclease and ligase. Extensive testing has demonstrated superior performance of BsaI-HFv2 compared to both BsaI (NEB #R0535) and BsaI-HF (NEB #R3535) in this challenging Golden Gate assembly context where restriction enzyme efficiency and fidelity are critically important.

High-Fidelity (HF®) restriction enzymes have the same specificity as native enzymes, but have been engineered for significantly reduced star activity and performance in a single buffer (rCutSmart™ Buffer).  All HF-restriction enzymes come with Gel Loading Dye, Purple (6X).  Enjoy the enhanced performance and added value of our engineered enzymes at the same price as the native enzyme:


  • Engineered for improved performance
  • Used in Golden Gate Assembly; maintains high activity levels in T4 DNA Ligase Buffer often used in this application
  • Type IIS restriction enzymes recognize asymmetric DNA sequences and cleave outside of their recognition sequence
  • 100% activity in rCutSmart Buffer
  • Time-Saver™ qualified for digestion in 5-15 minutes
  • Reduced star activity
  • NEB has developed convenient kits (using BsmBI-v2 and BsaI-HFv2) for performing Golden Gate Assembly.
  • Cut Site: GGTCTC(1/5)

"I really, really love BsaI-HFv2. We are back to getting phenomenal GGA results again with very little background."

    -M.C, Principal Investigator, Davidson College, NC


    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 rCutSmart™ Buffer
    Incubate at 37°C

    1X rCutSmart™ Buffer
    50 mM Potassium acetate
    20 mM Tris-acetate
    10 mM Magnesium acetate
    100 µg/ml Recombinant Albumin
    (pH 7.9 @ 25°C)

    Activity in NEBuffers

    NEBuffer™ r1.1: 100%
    NEBuffer™ r2.1: 100%
    NEBuffer™ r3.1: 100%
    rCutSmart™ Buffer: 100%

    Diluent Compatibility

    Storage Buffer

    10 mM Tris-HCl
    200 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: Impaired by Some Combinations of Overlapping
    CpG Methylation: Blocked by Some Combinations of Overlapping