Unit Definition

One unit is defined as the amount of enzyme required to digest 1 µg of λ 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

CutSmart® Buffer: 100%
NEBuffer 1.1: 10%
NEBuffer 2.1: 50%
NEBuffer 3.1: 10%

Diluent Compatibility

• Diluent B

Storage Conditions

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

Heat Inactivation

No

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Impaired by Overlapping

Companion Products

• 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

1. Cleavage of mammalian genomic DNA is impaired by some combinations of overlapping CpG methylation.

1. What does HF® refer to following the name of a restriction enzyme?
2. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
3. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
4. How can I access the old NEBuffer Activity Chart?
5. I tested your restriction enzyme on the substrate DNA recommended by NEB, and it appears to be active, however it does not digest my DNA. What could be the reason?
6. Is Gel Loading Dye, Purple (6X) or Gel Loading Dye, Purple (6X), no SDS compatible with other DNA binding dyes such as SYBR^® and GelRed™ during gel electrophoresis?
7. Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?
8. Why is my Restriction Enzyme not cutting DNA?
9. Why do I see additional DNA bands on my gel after a restriction digest?
10. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
11. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?

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

• Alphabetized List of Recognition Specificities
• Buffer and Diluent Formulation Table
• Compatible Cohesive Ends and Generation of New Restriction Sites
• Cross Index of Recognition Sequences
• Dam-Dcm and CpG Methylation
• Enzymes with Multiple Recognition Sequences
• Frequencies of Restriction Sites
• Interrupted Palindromes
• Isoschizomers
• Time-Saver™ Qualified Enzymes
• 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
• Dam and Dcm Methylases of E. coli
• Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
• Double Digests
• Effects of CpG Methylation on Restriction Enzyme Cleavage
• Heat Inactivation
• Megabase Mapping
• NEBuffer Activity/Performance Chart with Restriction Enzymes
• Optimizing Restriction Endonuclease Reactions
• Reduced Star Activities of HF® Enzymes
• 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 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]

• R3510S_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]

• R3510S_L_v1_0021303
• R3510S_L_v1_0021403
• R3510S_L_v1_0021309
• R3510S_L_v1_0021502
• R3510S_L_v1_0021507
• R3510S_L_v1_0031601
• R3510S_L_v1_0031606
• R3510S_L_v1_0031701
• R3510S_L_v1_0031704
• R3510S_L_v1_0031711

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

• DraIII-HF®
• Gel Loading Dye, Purple (6X)
• 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.