New England Biolabs
To access your account, log in or register.
Products Technical Reference Customer Service My NEB Account
Contact NEB About Us Site Map Request a Catalog OEM at NEB International Orders Freezer Program Quick Order
Related Information
FAQs for BspCNI
FAQs for Restriction Endonucleases
Technical Reference for Restriction Endonucleases
Favorite Tools
Enzyme Finder
NEBcutter
NEBuffer Chart
Double Digest Finder
PCR Selection Tool
Isoschizomers
DNA Sequences
and Maps
REBASE
Related Products
Reagents Sold Separately
NEBuffer 4
BSA
S-adenosylmethionine (SAM)
Discounts, Limited Offers and Value Purchases
BspCNI
NEBuffer 4BSASAM25Heat Inactivated
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:

CTCAG

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:
  1. The cleavage site of BspCNI varies.
  2. Two equally represented species of fragments are produced from BspCNI cleavage.

FAQs


  1. 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?
  2. Has the conversion to NEBuffer 4 altered any of the properties of the restriction enzyme?
  3. If I have an old tube of enzyme, what NEBuffer should I use?
  4. Will the new enzyme work in the originally supplied NEBuffer?
  5. 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
Privacy, Limitations, Warranty, Disclaimer, Copyright & Trademark