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FAQs for BspCNI FAQs for Restriction Endonucleases Technical Reference for Restriction Endonucleases Enzyme Finder NEBcutter NEBuffer Chart Double Digest Finder PCR Selection Tool Isoschizomers DNA Sequences and Maps REBASE NEBuffer 4 BSA S-adenosylmethionine (SAM)

Home > Products > Restriction Endonucleases > Restriction Endonucleases > BspCNI BspCNI Catalog # Size Concentration Price Qty   R0624L 500 units 2,000 units/ml $232.00 R0624S 100 units 2,000 units/ml $58.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF Recognition Site: isoschizomers | compatible ends | single letter code Source: Bacillus species (C. Nkenfou) Reagents Supplied: NEBuffer 4 (10X) BSA (100X) S-adenosylmethionine (SAM) (32 mM) Enzyme Properties Activity in NEBuffers: NEBuffer 1: 100% NEBuffer 2: 75% NEBuffer 3: 10% NEBuffer 4: 100% When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion. Methylation Sensitivity: dam methylation: Not sensitive dcm methylation: Not sensitive CpG methylation: Not sensitive More information about: Methylation Sensitivity Activity at 37°C: 75% Heat Inactivation: 80°C for 20 minutes Survival in a Reaction: (–) Not recommended for digest over 1 hour. More information about: Extended Digests with Restriction Enzymes Reaction & Storage Conditions Reaction Conditions: 1X NEBuffer 4 Supplemented with 100 μg/ml Bovine Serum Albumin and 20 μM S-adenosylmethionine (SAM) Incubate at 25°C. 1X NEBuffer 4: 20 mM Tris-acetate 50 mM potassium acetate 10 mM Magnesium Acetate 1 mM Dithiothreitol pH 7.9 @ 25°C Unit Definition: One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 25°C in a total reaction volume of 50 µl. Concentration: 2,000 units/ml Unit Assay Substrate: λ DNA Storage Conditions: 10 mM Tris-HCl 50 mM KCl 1 mM Dithiothreitol 0.1 mM EDTA 200 µg/ml BSA 50% Glycerol pH 7.4 @ 25°C Storage Temperature: -20°C Diluent Compatibility: Diluent A Notes General notes: The cleavage site of BspCNI varies. Two equally represented species of fragments are produced from BspCNI cleavage. FAQs The NEB catalog has historically stated that activity in NEBuffer 4 is less than 100% yet this enzyme is now supplied with NEBuffer 4. What have you changed? Has the conversion to NEBuffer 4 altered any of the properties of the restriction enzyme? If I have an old tube of enzyme, what NEBuffer should I use? Will the new enzyme work in the originally supplied NEBuffer? Why is NEB switching this restriction enzyme to NEBuffer 4? Quality Control for Current Lot Quality control values for a specific lot can be found on the datacard which accompanies each product. Ligation and Re-cutting: After a 5-fold overdigestion with BspCNI, approximately 50% of the DNA fragments can be ligated with T4 DNA Ligase (at a 5' termini concentration of 1-2 μM) at 16ºC. Of these ligated fragments, > 95% can be recut with BspCNI. 16-Hour Incubation: A 50 μl reaction containing 1 μg of DNA and 8 units of BspCNI incubated for 16 hours at 25ºC resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis. Exonuclease Activity: Incubation of a 50 μl reaction containing 4 units of BspCNI with 1 μg of a mixture of single and double-stranded [3H] E. coli DNA (205 cpm/μg) for 4 hours at 25ºC released < 0.2% of the total radioactivity. Reagents Sold Separately NEBuffer 4 BSA S-adenosylmethionine (SAM) New England Biolabs, Inc. is an ISO 9001 and ISO 14001 Certified Company. NEB certifies that it is a small business in accordance with the US Small Business Administration and 13 CFR 121.201