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.
An E. coli strain that carries the cloned BsaJI gene from Bacillus stearothermophilus J695 (Z. Chen)
The following reagents are supplied with this product:
Properties and Usage
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.
1X CutSmart™ Buffer Incubate at 37°C
1XCutSmart™ Buffer: 50 mM Potassium Acetate 20 mM Tris-acetate 10 mM Magnesium Acetate 100 μg/ml BSA pH 7.9 @ 25°C
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
65°C for 20 min
dam methylation: Not Sensitive dcm methylation: Blocked by Overlapping CpG Methylation: Blocked by Some Combinations of Overlapping
Activity at Temperature
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.
Blocked by overlapping dcm methylation. Cleavage of mammalian genomic DNA is blocked by some combinations of overlapping CpG methylation.
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%.
More information about: Methylation Sensitivity
Star activity may result from a glycerol concentration of >5%.
Legal and Disclaimers
New England Biolabs, Inc.
U.S. Patent No. 6,723,546
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 email@example.com.
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 firstname.lastname@example.org or fill out the Technical Support Form for appropriate document.