BsaI

This product was discontinued on 12/31/2020 and replaced with the High Fidelity version, BsaI-HFv2 (NEB #R3733).

Catalog # R0535 was discontinued on December 31, 2020

NEB restriction endonuclease that recognizes the sequence GGTCTCN^NNNN_

Isoschizomers | Single Letter Code | Pronunciation:

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^®) version available (NEB #R3733) supplied with CutSmart^®Buffer
Used in Golden Gate Assembly
Type IIS restriction enzymes recognize asymmetric DNA sequences and cleave outside of their recognition sequence
100% activity in CutSmart Buffer (over 210 enzymes are available in the same buffer) simplifying double digests
Supplied with 1 vial of Gel Loading Dye, Purple (6X)

Product Information
BsaI has a High Fidelity version BsaI-HF^®v2 (NEB #R3733).

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 BsaI gene from Bacillus stearothermophilus 6-55 (Z. Chen)

This product is related to the following categories:

Discontinued (<3 years)
- 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 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: 75%
  NEBuffer™ 3.1: 100%
  
  Diluent Compatibility
  
  Diluent B
  
  Storage Buffer
  
  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 minutes
  Methylation Sensitivity
  
  dam methylation: Not Sensitive
  dcm methylation: Impaired by Some Combinations of Overlapping
  CpG Methylation: Blocked by Some Combinations of Overlapping
- Related Products
  Companion Products
  
  dam^-/dcm^- Competent E. coli
  
  Monarch® Plasmid Miniprep Kit
  
  Monarch® DNA Gel Extraction Kit
  
  Monarch® PCR & DNA Cleanup Kit (5 μg)
  
  Materials Sold Separately
  
  Gel Loading Dye, Purple (6X)
  
  CutSmart® Buffer
- Product Notes
  Blocked by overlapping dcm methylation. Cleavage of mammalian genomic DNA is blocked by some combinations of overlapping CpG methylation.
  
  Activity at 50°C is 100%.
  
  May exhibit star activity in NEBuffer r1.1.
  
  Based on the stability of the enzyme in the reaction, incubations longerthan 1 hr will not result in improved digestion, unless additionalenzyme is added. Please refer to Restriction endonuclease survival in a reaction for more information regarding this topic.
  
  star activity may result from a glycerol concentration of >5%.
Protocols, Manuals & Usage
Tools & Resources
- Selection Charts
  Alphabetized List of Recognition Sequences
  
  Cleavage of Supercoiled DNA
  
  Compatible Cohesive Ends and Generation of New Restriction Sites
  
  Dam-Dcm and CpG Methylation
  
  Enzymes with Nonpalindromic Sequences
  
  Frequencies of Restriction Sites
  
  Isoelectric Points (pI) for Restriction Enzymes
  
  Isoschizomers
  
  NEB Diluent and Buffer Table
  
  Time-Saver™ Qualified Enzymes
  
  Type IIS Restriction Enzymes
  
  Why Choose Recombinant Enzymes?
- Web Tools
  Competitor Cross-Reference Tool
  
  DNA Sequences and Maps Tool
  
  Double Digest Finder
  
  Enzyme Finder
  
  NEBcutter™ v3.0
  
  NEBioCalculator®
  
  REBASE®
FAQs & Troubleshooting
- FAQs
  Which restriction enzymes are used in Golden Gate Assembly?
  
  Which restriction enzymes are used in GoldenBraid Assembly?
  
  Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
  
  What is the activity of the Type IIS restriction enzyme BsaI-HFv2 (NEB #R3733) in T4 DNA Ligase Buffer?
  
  Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  
  Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?
- Troubleshooting
  Restriction Enzyme Troubleshooting Guide
- Tech Tips
  
  Restriction Enzymes in Golden Gate Assembly
Citations & Technical Literature
- Brochures
  Golden Gate Assembly Brochure
- Citations
  Product Citation Tool
  
  Powered by Bioz See more details on Bioz
  
  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
  
  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
  
  Sakuma T, Nishikawa A, Kume S, Chayama K, Yamamoto T (2014) Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system Sci Rep; 4, 5400. PubMedID: 24954249, DOI: 10.1038/srep05400
Quality, Safety & Legal
- Quality Control Assays
  
  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.
- Specifications & Change Notifications
  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]
  
  R0535S_L_v2
- 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]
  
  R0535S_L_v1_0371403
  
  R0535S_L_v1_0361212
  
  R0535S_L_v1_0361305
  
  R0535S_L_v1_0361311
  
  R0535S_L_v1_0371404
  
  R0535S_L_v2_0371404
  
  R0535S_L_v1_0371409
  
  R0535S_L_v2_0371503
  
  R0535S_L_v2_0371509
  
  R0535S_L_v2_0371603
  
  R0535S_L_v2_0381608
  
  R0535S_L_v2_0401702
  
  R0535S_L_v2_0401708
  
  R0535S_L_v2_0431712
  
  R0535S_L_v2_0431805
  
  R0535L_v2_10009918
  
  R0535S_v2_10018570
  
  R0535L_v2_10030542
  
  R0535S_v2_10030544
  
  R0535S_v2_10043185
  
  R0535L_v2_10043183
  
  R0535S_v2_10055060
  
  R0535S_v2_10059599
  
  R0535S_v2_10062820
  
  R0535L_v2_10062822
  
  R0535S_v2_10065817
  
  R0535S_v2_10081423
  
  R0535S_v2_10091140
  
  R0535L_v2_10086217
- Safety Data Sheets
  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
  
  BsaI
- Legal and Disclaimers
  
  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.