High-Fidelity (HF®) Restriction Endonucleases

Restriction Endonucleases
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  • For over 40 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 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.

    EcoRI-HF® (NEB #R3101) shows no star activity in overnight digests, even when used at higher concentrations. 50 μl reactions were set up using 1 μg of Lambda DNA, the indicated amount of enzyme and the recommended reaction buffer. Reactions were incubated overnight at 37°C. Marker M is the 1 kb DNA Ladder (NEB# N3232)

    HF® is a registered trademark of New England Biolabs, Inc.
    FastDigest® is a registered trademark of Fermentas.

    Benefits of High-Fidelity (HF®) Restriction Enzymes

    • 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


    • Cloning
    • Genotyping
    • Mutational Analysis
    • Mapping
    • Probe Preparation 
    • Sequencing 
    • Methylation Detection
    • Any application that requires high fidelity or flexible reaction setup