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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Version 1.0–02.12
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