High-Fidelity (HF®) Restriction Endonucleases

Restriction Endonucleases

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

Applications

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