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BsaI-HF®v2

Catalog # Concentration Size List Price Quantity Your Price
R3733G 20000 units/ml 300 units
Please Inquire
R3733L 20000 units/ml 5000 units $468.00
$421.20
R3733S 20000 units/ml 1000 units $112.00
$100.80
Catalog # Size List Price Your Price
R3733G 300 units
R3733L 5000 units $468.00
$421.20
R3733S 1000 units $112.00
$100.80
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
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
  • Optimized for 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.
  • Can be used with NEBridge® Ligase Master Mix for Golden Gate Assembly
  • Restriction Enzyme 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

Featured Videos
View Video Library
  • Monarch_MiniprepProtocol_VideoThumb
    Golden Gate Assembly Workflow
  • Monarch_MiniprepProtocol_VideoThumb
    Golden Gate Assembly Domestication Tutorial
  • Monarch_MiniprepProtocol_VideoThumb
    Listen to DAD Informatics tools and NEB enzymes to enable complex one-pot Golden Gate Assemblies
  • Monarch_MiniprepProtocol_VideoThumb
    Listen to DAD (Data-optimized Assembly Design) when constructing high-complexity Golden Gate Assembly targets
  • Monarch_MiniprepProtocol_VideoThumb
    Golden Gate Assembly Tool Tutorial
  • Monarch_MiniprepProtocol_VideoThumb
    Restriction Enzymes in Golden Gate Assembly
  • Monarch_MiniprepProtocol_VideoThumb
    Function and Application of Type IIS Restriction Enzymes

BsaI-HF®v2 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′.

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 and modified BsaI gene from Bacillus stearothermophilus 6-55 (Z. Chen)
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  BsaI-HF®v2 R3733SVIAL -20 1 x 0.05 ml 20,000 units/ml
  Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X
  rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X
  BsaI-HF®v2 R3733LVIAL -20 1 x 0.25 ml 20,000 units/ml
  rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X
  Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X

Properties & 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 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

NEBufferâ„¢ r1.1: 100%
NEBufferâ„¢ r2.1: 100%
NEBufferâ„¢ r3.1: 100%
rCutSmartâ„¢ Buffer: 100%

Diluent Compatibility

Storage Buffer

20 mM Tris-HCl
300 mM NaCl
0.1 mM TCEP
200 µg/ml Recombinant Albumin
50% Glycerol
pH 9 @ 25°C

Heat Inactivation

80°C for 20 minutes

Methylation Sensitivity

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



Companion Products
Notes
  • BsaI-HFv2 has the same specificity as BsaI (NEB #R0535), but it has been engineered for reduced star activity.
  • Impaired by some combinations of overlapping dcm methylation and blocked by some combinations of overlapping CpG methylation.
FAQs
Troubleshooting
Citations
Additional Citations
  • Lee JH, Won HJ, Oh E-S, Oh M-H and Jung JH (2020) Golden Gate Cloning-Compatible DNA Replicon/2A-Mediated Polycistronic Vectors for Plants Front Plant Sci 11, 559365. DOI: 10.3389/fpls.2020.559365
Publications
  • Thuronyi, B.W., Koblan, L.W., Levy, J.M. et al (2019). Continuous evolution of base editors with expanded target compatibility and improved activity Nat Biotechnol. 37, 1070-1079.. DOI: 10.1038/s41587-019-0193-0
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Change Notifications
BsaI-HFv2 has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10156743. All subsequent (higher number) lots will contain rAlbumin. For additional information, please visit at NEB Restriction Enzyme formulations with Recombinant Albumin (rAlbumin).
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]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
Legal And Disclaimer

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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