High-Fidelity (HF®) Restriction Endonucleases
For over 45 years, New England Biolabs has been developing innovative solutions for molecular biology applications. The respected leader in the field of restriction enzyme biology, NEB has developed a line of High-Fidelity (HF®) Restriction Enzymes. These engineered enzymes have the same specificity as the native enzyme, with the added benefit of reduced star activity, rapid digestion (5-15 minutes) and 100% activity in rCutSmart™ and CutSmart® Buffer.
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.
Optimized performance for a wide range of conditions
High Fidelity Restriction Enzymes have been engineered by exchanging functional amino acid residues and then screening for optimal mutants that perform under a wide range of conditions. Whether you are setting up digests for 5-15 minutes or overnight, or using varying amounts of enzymes, HF enzymes ensure the performance you need.
We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.
HF® is a registered trademark of New England Biolabs, Inc.
FastDigest® is a registered trademark of Fermentas.
Restriction Enzymes at NEB: Over 30 years of Innovation
A Modern Day Gene Genie Sir Richard Roberts on Rebase
- Competitor Cross-Reference Tools
- DNA Sequences and Maps Tool
- Enzyme Finder
- Alphabetized List of Recognition Sequences
- Cleavage Of Supercoiled DNA
- Compatible Cohesive Ends and Generation of New Restriction Sites
- Dam-Dcm and CpG Methylation
- Enzymes with Multiple Recognition Sequences
- Enzymes with Nonpalindromic Sequences
- Frequencies of Restriction Sites
- Interrupted Palindromes
- Recleavable Blunt Ends
- Recleavable Filled-in 5' Overhangs
- Restriction Enzyme Troubleshooting Guide
- Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
- Activity of Restriction Enzymes in PCR Buffers
- Alteration of Apparent Recognition Specificities Using Methylases
- 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
- Heat Inactivation
- 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
- Restriction of Foreign DNA by E. coli K-12
- Site Preferences
- Star Activity
- Kamps-Hughes, N., Quimby, A., Zhu, Z., Johnson, E.A. (2013) Massively parallel characterization of restriction endonucleases Nucleic Acids Res ; 41(11), e119. PubMedID: 23605040, DOI: 10.1093/nar/gkt257
- One buffer convenience with no loss of performance
- Reduced star activity eliminates unwanted cleavage
- Time-Saver qualified for 5-15 minute digests, flexible enough to digest overnight.
- Engineered for performance under a wide range of conditions
- Added flexibility without added cost
- Mutational Analysis
- Probe Preparation
- Methylation Detection
- Any application that requires high fidelity or flexible reaction setup
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email firstname.lastname@example.org.
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.
Reduce Star Activity with High-Fidelity Restriction Enzymes
NEB has engineered HF® enzymes to eliminate star activity. Learn how, and what this means for your digests.
Discovering New Restriction Enzymes
Watch as Rick Morgan, Research Scientist in the Restriction Enzyme Division, describes his passion for discovering and characterizing restriction enzymes from nature.
How do restriction enzymes interact with DNA?
Watch as Geoff Wilson, Restriction Enzyme Division Head, describes the interaction of restriction enzymes and substrate DNA using computer models generated from x-ray crystallography data.
Restriction Enzyme Digest Problem: Too Many DNA Bands
Are you finding unexpected bands in your digestion reaction? Here are some tips to help you determine the cause.
What is Restriction Enzyme Star Activity?
Learn what Star Activity is, why it is detrimental to accurate restriction enzyme digestion, and how NEB's HF enzymes are engineered to avoid it.
NEB® Restriction Enzyme Double Digest Protocol
Double digestions can save you time, and this video can offer tips for how to achieve the best results, no matter which of NEB's restriction enzymes you're using.
Restriction Enzyme Digestion Problem: DNA Smear on Agarose Gel
Learn more about what causes this common problem, and how NEB's enzymes are QC'd to avoid DNA smearing.
Standard Protocol for Restriction Enzyme Digests
Let one of NEB's restriction enzyme experts help you improve your technique and avoid common mistakes in digest setup.