Overview            Features             Applications   

Engineered Enzymes with Increased Fidelity and Buffer Compatibility
As part of our ongoing commitment to the study and improvement of restriction enzymes, NEB has made several advances in restriction enzyme research. This work has resulted in innovative new products, including a line of High Fidelity (HF) restriction enzymes. These engineered enzymes have the same specificity as their established counterpart with the benefit of reduced star activity. Experience a new level of flexibility in reaction conditions, including:

• Volume choice
• Fast digestion times (5-15 minutes)
• All work in the same buffer

	Engineered for performance
	Dramatically reduced star activity (up to 500-fold in some cases)
	100% Activity in a single buffer
	Time-Saver™ qualified for 5-15 minute digests
	Value – Same price as established enzymes

	Features: Reduced star activity Single buffer (NEBuffer 4) Time-Saver qualified (digests substrate DNA in 5-15 minutes) Added flexibility in digest setup No added expense – available at same low price per unit as wild type enzyme
EcoRI-HF™ shows significantly reduced star activity, as compared to other enzymes that digest DNA in 5-15 minutes. EcoRI-HF shows no star activity when used in 5 minutes or overnight. 50 μl reactions were set up using 1 μg of Lambda DNA, 1 μl of enzyme and recommended reaction buffer. Digests were incubated at 37°C. Marker M is the 1kb DNA Ladder (NEB# N3232).	Applications: Cloning Genotyping Mutational Analysis Mapping Probe Preparation Sequencing Methylation Detection Any application that requires high fidelity or flexible reaction setup
EcoRI-HF 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 at 37°C. Marker M is the 1kb DNA Ladder (NEB# N3232).