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 eliminating offtarget products, while offering convenience and flexibility in experimental design. Benefits of High Fidelity (HF) Restriction Enzymes: 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

3 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, 1 hour 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 New 203 restriction enzymes 100% active in NEBuffer 4 England Biolabs currently supplies 279 restriction enzymes, 203 of which are 100% 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.

4 What does an HF Restricti Engineered for performance 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 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 100% activity 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! Faster digests with the same reliable 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 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 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.

5 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 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. For full ordering information, including prices, extensive technical information, protocols etc., please visit Indicates a RE-Mix Master Mix format is available. ProdUct RECOGNITION SITE NEB # BUffEr Hf factor AgeI-HF 5... A C C G G T... 3 R3552S/L 4 + BSA 8 AgeI 3... T G G C C A... 5 R0552S/L 1 BamHI-HF 5... G G A T C C... 3 R3136S/L BamHI 3... C C T A G G... 5 R0136S/L 3 + BSA BmtI-HF 5... G C T A G C... 3 R3658S/L 4 + BSA 62,500 BmtI 3... C G A T C G... 5 R0658S/L 2 BsaI-HF 5... G G T C T C (N) R3535S/L 4 + BSA 250 BsaI 3... C C A G A G (N) R0535S/L 4 + BSA BstEII-HF R3162S/L 4 + BSA G G T N A C C... 3 BstEII 3... C C A N T G G... 5 R0162S/L 3 + BSA DraIII-HF 5... C A C N N N G T G... 3 R3510S/L 4 1,000 DraIII 3... G T G N N N C A C... 5 R0510S/L 3 + BSA EagI-HF 5... C G G C C G... 3 R3505S/L 4 2 EagI 3... G C C G G C... 5 R0505S/L 3 EcoRI-HF 5... G A A T T C... 3 R3101S/L 4 64 EcoRI 3... C T T A A G... 5 R0101S/L U EcoRV-HF 5... G A T A T C... 3 R3195S/L 4 64 EcoRV 3... C T A T A G... 5 R0195S/L 3 + BSA HindIII-HF 5... A A G C T T... 3 R3104S/L 4 2,000 HindIII 3... T T C G A A... 5 R0104S/L 2 KpnI-HF 5... G G T A C C... 3 R3142S/L 4 62,500 KpnI 3... C C A T G G... 5 R0142S/L 1 + BSA MfeI-HF 5... C A A T T G... 3 R3589S/L 4 16 MfeI 3... G T T A A C... 5 R0589S/L 4 NcoI-HF R3193S/L C C A T G G... 3 NcoI 3... G G T A C C... 5 R0193S/L 3 NheI-HF 5... G C T A G C... 3 R3131S/L 4 + BSA 266 NheI 3... C G A T C G... 5 R0131S/L 2 + BSA NotI-HF 5... G C G G C C G C... 3 R3189S/L 4 + BSA 16 NotI 3... C G C C G G C G... 5 R0189S/L 3 + BSA PstI-HF 5... C T G C A G... 3 R3140S/L 4 33 PstI 3... G A C G T C... 5 R0140S/L 3 + BSA PvuI-HF 5... C G A T C G... 3 R3150S/L 4 + BSA 32 PvuI 3... G C T A G C... 5 R0150S/L 3 + BSA PvuII-HF 5... C A G C T G... 3 R3151S/L 4 32 PvuII 3... G T C G A C... 5 R0151S/L 2 SacI-HF 5... G A G C T C... 3 R3156S/L 4 + BSA 266 SacI 3... C T C G A G... 5 R0156S/L 1 + BSA SalI-HF R3138S/L 4 8, G T C G A C... 3 SalI 3... C A G C T G... 5 R0138S/L 3 + BSA SbfI-HF 5... C C T G C A G G... 3 R3642S/L 4 32 SbfI 3... G G A C G T C C... 5 R0642S/L 4 ScaI-HF 5... A G T A C T... 3 R3122S/L 4 62 ScaI 3... T C A T G A... 5 R0122S/L 3 SpeI-HF 5... A C T A G T... 3 R3133S/L 4 + BSA 16 SpeI 3... T G A T C A... 5 R0133S/L 4 + BSA SphI-HF 5... G C A T G C... 3 R3182S/L SphI 3... C G T A C G... 5 R0182S/L 2 SspI-HF 5... A A T A A T... 3 R3132S/L 4 16 SspI 3... T A A T A A... 5 R0132S/L U StyI-HF 5... C C W W G G... 3 R3500S/L StyI 3... G G W W C C... 5 R0500S/L 3 + BSA

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