Unit Definition

One unit is defined as the amount of enzyme required to convert 1 µg of supercoiled pUC19 DNA to open circular form in 1 hour at 65°C in a total reaction volume of 50 µl.

Reaction Conditions

IX CutSmart® Buffer
Incubate at 65°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

CutSmart® Buffer: 100%
NEBuffer 1.1: 25%
NEBuffer 2.1: 100%
NEBuffer 3.1: 100%

Diluent Compatibility

• Diluent A

Storage Temperature

-20°C

Storage Conditions

0.1 mM EDTA
50% Glycerol
1 mM DTT
10 mM Tris-HCl
200 μg/ml BSA
200 mM NaCl
pH 7.4 @ 25°C

Heat Inactivation

80°C for 20 min

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive

Activity at Temperature

@37°C: 75%

Companion Products

• Monarch® Plasmid Miniprep Kit 
• Monarch® DNA Gel Extraction Kit
• Monarch® PCR & DNA Cleanup Kit (5 μg)

Materials Sold Separately

• CutSmart® Buffer

1. To see how this enzyme (along with other nicking enzymes) performs at different temperatures, please visit Effect of Various Temperatures of Different Nicking Endonucleases.

1. Song, Q. et al. (2010). Anal. Chem. [Epub ahead of print].
2. Zhang, P. et al. (2010). Protein Expr. Purif. 69, 226-234. [Epub 2009 Sep 9].

1. My enzyme used to come with another NEBuffer, but CutSmart™ Buffer is now recommended?  Why?
2. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
3. How can I access the old NEBuffer Activity Chart?
4. Why is my Restriction Enzyme not cutting DNA?
5. Why do I see a DNA smear on an agarose gel after a restriction digest?
6. Why do I see additional DNA bands on my gel after a restriction digest?
7. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?

1. Optimizing Restriction Endonuclease Reactions
2. Double Digest Protocol with Standard Restriction Enzymes

• Alphabetized List of Recognition Specificities
• Buffer and Diluent Formulation Table
• Cross Index of Recognition Sequences
• Dam-Dcm and CpG Methylation
• Effect of Various Temperatures on Nicking Endonucleases
• Enzymes with Nonpalindromic Sequences
• Frequencies of Restriction Sites
• Isoschizomers
• Why Choose Recombinant Enzymes?

• Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
• Activity of Restriction Enzymes in PCR Buffers
• Cleavage Close to the End of DNA Fragments
• Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
• Double Digests
• Heat Inactivation
• Megabase Mapping
• NEBuffer Activity/Performance Chart with Restriction Enzymes
• Optimizing Restriction Endonuclease Reactions
• Restriction Endonucleases - Survival in a Reaction
• Restriction Enzyme Diluent Buffer Compatibility
• Restriction Enzyme Tips
• Single Letter Codes
• Star Activity
• Traditional Cloning Quick Guide

• Restriction Enzyme Troubleshooting Guide

• DNA Sequences and Maps Tool

• Double Digest Finder
• Enzyme Finder
• NEBcutter V2.0
• NEBioCalculator
• REBASE

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.

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]

• R0648S_L_v1

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

• R0648S_L_v1_0041302
• R0648S_L_v1_0041312
• R0648S_L_v1_0041408
• R0648S_L_v1_0041503
• R0648S_L_v1_0041507
• R0648S_L_v1_0041601
• R0648S_L_v1_0041602
• R0648S_L_v1_0041606
• R0648S_L_v1_0041611

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• Nb.BsrDI
• CutSmart® Buffer

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at gbd@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.