High Fidelity (HF) Restriction Enzymes

Transcription

1 High Fidelity (HF) Restriction Enzymes

2 Performance you can count on High Fidelity (HF ) Restriction Enzymes from New England Biolabs 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 fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design. 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 Engineered for performance under a wide range of conditions Added flexibility without added cost

3 What does an HF Restricti 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 optimum mutants that perform under a wide range of conditions. Whether you are setting up digests for 5 minutes or overnight, or using varying amounts of enzyme, HF enzymes ensure the performance you need. Flexible enzyme input with reliable results NEB EcoRI-HF FastDigest EcoRI M µl 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). One-buffer convenience with no loss of 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 180 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! Faster digests with the same great results 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 withstand overnight digestions. Choose the reaction time that works for you. For a complete list of Time-Saver qualified enzymes, visit Predictable performance in 5 minutes or overnight NEB EcoRI-HF FastDigest EcoRI o/n 5 min M o/n 5 min 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). HF is a trademark of New England Biolabs, Inc. FastDigest is a registered trademark of Fermentas.

4 on Enzyme mean to you? Greater confidence in the accuracy of your results Star activity, an intrinsic property of restriction enzymes, describes the unwanted cleavage and end-product degradation that occurs under suboptimal reaction conditions. When used with an incompatible buffer or at a high enzyme concentration, offtarget 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 number of units of HF enzyme that can be used compared to the wild type before significant star activity is detected. The HF factor refers to the X-fold reduction in star activity that is achieved by choosing an HF enzyme, and clearly illustrates the flexibility that is offered by using an HF enzyme. For full ordering information, visit Indicates a RE-Mix Master Mix format is available. PRODUCT RECOGNITION SITE NEB # BUFFER MAXIMUM UNITS WITH NO STAR ACTIVITY HF FACTOR AgeI-HF 5... A C C G G T... 3 R3552S/L 4 + BSA AgeI 3... T G G C C A... 5 R BamHI-HF 5... G G A T C C... 3 R3136S/L 4 4, BamHI 3... C C T A G G... 5 R BSA 32 BmtI-HF 5... G C T A G C... 3 R BSA 1,000,000 62,500 BmtI 3... C G A T C G... 5 R BsaI-HF 5... G G T C T C (N) R3535S/L 4 + BSA 8, BsaI 3... C C A G A G (N) R BSA 32 BstEII-HF 5... G G T N A C C... 3 R BSA 2, BstEII 3... C C A N T G G... 5 R BSA 16 DraIII-HF 5... C A C N N N G T G... 3 R3510S/L 4 2,000 1,000 DraIII 3... G T G N N N C A C... 5 R BSA 2 EagI-HF 5... C G G C C G... 3 R3505S/L EagI 3... G C C G G C... 5 R EcoRI-HF R3101S/L 4 16, G A A T T C... 3 EcoRI 3... C T T A A G... 5 R0101 U 250 EcoRV-HF 5... G A T A T C... 3 R3195S/L 4 64, EcoRV 3... C T A T A G... 5 R BSA 1,000 HindIII-HF 5... A A G C T T... 3 R3104S/L 4 500,000 2,000 HindIII 3... T T C G A A... 5 R KpnI-HF 5... G G T A C C... 3 R3142S/L 4 1,000,000 62,500 KpnI 3... C C A T G G... 5 R BSA 16 MfeI-HF 5... C A A T T G... 3 R3589S/L MfeI 3... G T T A A C... 5 R NcoI-HF 5... C C A T G G... 3 R3193S/L 4 64, NcoI 3... G G T A C C... 5 R NheI-HF R3131S/L 4 + BSA 32, G C T A G C... 3 NheI 3... C G A T C G... 5 R BSA 12 NotI-HF 5... G C G G C C G C... 3 R3189S/L 4 + BSA 64, NotI 3... C G C C G G C G... 5 R BSA 4,000 PstI-HF 5... C T G C A G... 3 R3140S/L 4 4, PstI 3... G A C G T C... 5 R BSA 120 PvuI-HF 5... C G A T C G... 3 R3150S/L 4 + BSA 16, PvuI 3... G C T A G C... 5 R BSA 500 PvuII-HF 5... C A G C T G... 3 R3151S/L PvuII 3... G T C G A C... 5 R SacI-HF 5... G A G C T C... 3 R3156S/L 4 + BSA 32, SacI 3... C T C G A G... 5 R BSA 120 SalI-HF R3138S/L 4 32,000 8, G T C G A C... 3 SalI 3... C A G C T G... 5 R BSA 4 SbfI-HF 5... C C T G C A G G... 3 R3642S/L SbfI 3... G G A C G T C C... 5 R ScaI-HF 5... A G T A C T... 3 R3122S/L ScaI 3... T C A T G A... 5 R SpeI-HF 5... A C T A G T... 3 R3133S/L 4 + BSA 8, SpeI 3... T G A T C A... 5 R BSA 500 SphI-HF 5... G C A T G C... 3 R3182S/L 4 8, SphI 3... C G T A C G... 5 R SspI-HF 5... A A T A A T... 3 R3132S/L SspI 3... T A A T A A... 5 R0132 U 32 StyI-HF 5... C C W W G G... 3 R3500S/L 4 4, StyI 3... G G W W C C... 5 R BSA 32

5 Convenience you are looking for Single buffer simplicity 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 or want to leave your reactions overnight, restriction enzyme digests with HF enzymes produce the same, reliable results. Set up your digests with reaction-specific enzyme concentrations, without fear of risking product integrity to star activity. HF enzymes offer added flexibility and functionality without added cost they re available for the same price-per-unit as the standard version. HF restriction enzymes from NEB part of the largest single buffer restriction enzyme collection 279 New 182 restriction enzymes active in NEBuffer 4 England Biolabs currently supplies 279 restriction enzymes, 182 of which are active in NEBuffer 4. Mix it up. For added convenience, many of NEB s HF enzymes are available in RE-Mix Master Mix format. Visit for more details.

6 USA New England Biolabs, Inc. 240 County Road Ipswich, MA France New England Biolabs France Telephone : info@fr.neb.com Telephone: (978) Toll Free: (U.S. Orders) Toll Free: (U.S. Tech) Fax: (978) info@neb.com Germany New England Biolabs GmbH Free Call: 0800/ info@de.neb.com Canada New England Biolabs, Ltd. Toll Free: info@ca.neb.com China, People s Republic New England Biolabs (Beijing), Ltd. Telephone: / info@neb-china.com Japan New England Biolabs Japan, Inc. Telephone: +81 (0) info@neb-japan.com United Kingdom New England Biolabs (UK), Ltd. Call Free info@uk.neb.com Accessibility Mobile tools for your research Now available for the Droidtm and iphone : NEBTools to aid in setting up your restriction enzyme digests. DROID is a trademark of Lucasfilm, Ltd. and its related companies. Used under license. iphone is a registered trademark of Apple Inc. Version