High Fidelity (HF) Restriction Enzymes

For over 35 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 High Fidelity (HF) Restriction Enzymes to address two short-comings of traditional restriction enzymes: buffer compatibility and star activity.

HF restriction enzymes have 100% activity in NEBuffer 4; single-buffer simplicity means more straightforward and streamlined sample processing. HF enzymes also exhibit dramatically reduced star activity. Engineered with performance in mind, HF restriction enzymes are eliminating off-target products, while offering convenience and flexibility in experimental

design.

High Fidelity (HF™) Restriction Enzymesfrom New England Biolabs

Performance you can count on

One buffer convenience with 100% activity

Dramatically reduced star activity eliminates unwanted cleavage

Time-Saver qualified for 5–15-minute digests

Engineered for performance

Added flexibility without added cost

Benefits of High Fidelity (HF) Restriction Enzymes:

Gone are the days of buffer compatibility analyses and sub-optimal enzyme conditions! HF restriction enzymes are all 100% active in a single buffer: NEBuffer 4. This simplifies double digest reactions and eliminates the need for setting up sequential digests. HF enzymes share the same site specificity and cleavage efficiency as their wild-type counterparts, but offer the increased flexibility and convenience that you need in your laboratory. Whether you have 5 minutes, 1 hour or want to leave your reactions over-night, restriction enzyme digests with HF enzymes produce the same, reli-able results. Set up your digests with reaction-specific enzyme concentra-tions, without fear of risking product integrity to star activity. HF enzymes offer added flexibility and functionality without added cost – they’re avail-able for the same price-per-unit as the standard version.

Convenience you are looking forSingle buffer simplicity

Mix it up.For added convenience, many of NEB’s HF enzymes are available in RE-Mix™ Master Mix format.

Visit www.neb.com for more details.

HF restriction enzymes from NEB – part of thelargest single buffer restriction enzyme collection

203 restriction enzymes 100% active in NEBuffer 4

279279279279279279279279279279279279279 New England Biolabs currently supplies 279 restriction enzymes, 203 of which are 100% active in NEBuffer 4.

What does an HF Restriction Enzyme mean to you?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 overnight at 37°C. Marker M is the 1 kb DNA Ladder (NEB# N3232).

1 3 5 10 M 1 3 5 10 µl

NEB EcoRI-HF™ FastDigest® EcoRI

Flexible enzyme input with reliable results

High Fidelity Restriction Enzymes have been engineered by exchanging functional amino acid residues and then stringently screened for optimum mutants: Whether you are setting up digests for 5 minutes or overnight, or using varying amounts of enzyme, HF enzymes ensure the perfor-mance you need.

Engineered for performance

All High Fidelity Restriction Enzymes work optimally in NEBuffer 4, which offers the highest level of compatibility in the NEBuffer System. In fact, over 200 restriction enzymes exhibit 100% activity in NEBuffer 4, which is the largest number of enzymes commercially available for use with a single buffer. No more checking for buffer compatability!

One-buffer convenience with 100% activity

All HF enzymes are Time-Saver qualified and certified to be nuclease free, giving them the power to digest in 5-15 minutes and the purity to safely set up digests for long periods of time without any degradation of your sample. Choose the reaction time that works for you. For a complete list of Time-Saver qualified enzymes, visit www.neb.com

Faster digests with the same reliable results

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 1 kb DNA Ladder (NEB# N3232).

Predictable performance in 5 minutes or overnight

o/n 5 min M o/n 5 min

NEB EcoRI-HF™ FastDigest® EcoRI

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

What does an HF Restriction Enzyme mean to you?

Star activity, an intrinsic property of restriction enzymes, describes the unwanted cleavage and end-product degradation that occurs under sub-optimal reaction conditions. When used with an incompatible buffer or at a high enzyme concentration, off-target and degenerate products can be formed. HF enzymes have been engineered for reduced star activity, increasing accuracy and avoiding unwanted cleavage.

The table indicates the HF factor that refers to the x-fold reduction in star activity that is achieved by choosing an HF enzyme in relation to their original prototypes, and clearly illustrates the benefit that is offered by using an HF enzyme.

Greater confidence in the accuracy of your results

* Indicates a RE-Mix Master Mix format is available.

For full ordering information, including prices, extensive technical information, protocols etc., please visit www.neb.com

ProdUct RECOGNITION SITE NEB # BUffEr Hf fActor

AgeI-HF™

* R3552S/L 4 + BSA ≥ 8

AgeI R0552S/L 1

BamHI-HF™ R3136S/L 4 ≥ 125

BamHI R0136S/L 3 + BSA

BmtI-HF™ R3658S/L 4 + BSA 62,500

BmtI R0658S/L 2

BsaI-HF™ R3535S/L 4 + BSA ≥ 250

BsaI R0535S/L 4 + BSA

BstEII-HF™

* R3162S/L 4 + BSA ≥ 125

BstEII R0162S/L 3 + BSA

DraIII-HF™ R3510S/L 4 ≥ 1,000

DraIII R0510S/L 3 + BSA

EagI-HF™ R3505S/L 4 2

EagI R0505S/L 3

EcoRI-HF™

* R3101S/L 4 64

EcoRI R0101S/L U

EcoRV-HF™

* R3195S/L 4 ≥ 64

EcoRV R0195S/L 3 + BSA

HindIII-HF™ R3104S/L 4 ≥ 2,000

HindIII R0104S/L 2

KpnI-HF™

* R3142S/L 4 ≥ 62,500

KpnI R0142S/L 1 + BSA

MfeI-HF™

* R3589S/L 4 ≥ 16

MfeI R0589S/L 4

NcoI-HF™

* R3193S/L 4 ≥ 530

NcoI R0193S/L 3

NheI-HF™

* R3131S/L 4 + BSA ≥ 266

NheI R0131S/L 2 + BSA

NotI-HF™ R3189S/L 4 + BSA ≥ 16

NotI R0189S/L 3 + BSA

PstI-HF™ R3140S/L 4 33

PstI R0140S/L 3 + BSA

PvuI-HF™ R3150S/L 4 + BSA ≥ 32

PvuI R0150S/L 3 + BSA

PvuII-HF™ R3151S/L 4 32

PvuII R0151S/L 2

SacI-HF™ R3156S/L 4 + BSA ≥ 266

SacI R0156S/L 1 + BSA

SalI-HF™

* R3138S/L 4 ≥ 8,000

SalI R0138S/L 3 + BSA

SbfI-HF™ R3642S/L 4 32

SbfI R0642S/L 4

ScaI-HF™

* R3122S/L 4 62

ScaI R0122S/L 3

SpeI-HF™ R3133S/L 4 + BSA ≥ 16

SpeI* R0133S/L 4 + BSA

SphI-HF™ R3182S/L 4 250

SphI R0182S/L 2

SspI-HF™ R3132S/L 4 16

SspI R0132S/L U

StyI-HF™ R3500S/L 4 125

StyI R0500S/L 3 + BSA

5´. . . A C C G G T . . . 3´

3´. . . T G G C C A . . . 5´

5´. . . C C W W G G . . . 3´

3´. . . G G W W C C . . . 5´

5´. . . A A T A A T . . . 3´

3´. . . T A A T A A . . . 5´

5´. . . G C A T G C . . . 3´

3´. . . C G T A C G . . . 5´

5´. . . A G T A C T . . . 3´

3´. . . T C A T G A . . . 5´

5´. . . C C T G C A G G . . . 3´

3´. . . G G A C G T C C . . . 5´

5´. . . G T C G A C . . . 3´

3´. . . C A G C T G . . . 5´

5´. . . G A G C T C . . . 3´

3´. . . C T C G A G . . . 5´

5´. . . C A G C T G . . . 3´

3´. . . G T C G A C . . . 5´

5´. . . C G A T C G . . . 3´3´. . . G C T A G C . . . 5´

5´. . . C T G C A G . . . 3´

3´. . . G A C G T C . . . 5´

5´. . . G C G G C C G C . . . 3´

3´. . . C G C C G G C G . . . 5´

5´. . . G C T A G C . . . 3´

3´. . . C G A T C G . . . 5´

5´. . . C C A T G G . . . 3´

3´. . . G G T A C C . . . 5´

5´. . . C A A T T G . . . 3´

3´. . . G T T A A C . . . 5´

5´. . . G G T A C C . . . 3´

3´. . . C C A T G G . . . 5´

5´. . . A A G C T T . . . 3´3´. . . T T C G A A . . . 5´

5´. . . G A T A T C . . . 3´

3´. . . C T A T A G . . . 5´

5´. . . G A A T T C . . . 3´

3´. . . C T T A A G . . . 5´

5´. . . C G G C C G . . . 3´

3´. . . G C C G G C . . . 5´

5´. . . C A C N N N G T G . . . 3´3´. . . G T G N N N C A C . . . 5´

5´. . . G G T C T C (N)1 . . . 3´

3´. . . C C A G A G (N)5 . . . 5´

5´. . . G G A T C C . . . 3´

3´. . . C C T A G G . . . 5´

5´. . . G C T A G C . . . 3´3´. . . C G A T C G . . . 5´

5´. . . A C T A G T . . . 3´

3´. . . T G A T C A . . . 5´

5´. . . G G T N A C C . . . 3´

3´. . . C C A N T G G . . . 5´

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AccessibilityMobile tools for your research

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