Fidelity at its Finest.
Q5® High-Fidelity DNA Polymerase (NEB #M0491) sets a new standard for both fidelity and robust performance. With the highest fidelity amplification available (>280 times higher than Taq), Q5 DNA Polymerase results in ultra-low error rates. Q5 DNA Polymerase is composed of a novel polymerase that is fused to the processivity-enhancing Sso7d DNA binding domain, improving speed, fidelity and reliability of performance.
NEW: Q5U Hot Start High-Fidelity DNA Polymerase (NEB #M0515). Q5U is a modified version of Q5 High-Fidelity DNA Polymerase containing a mutation in the uracil-binding pocket that enables the ability to read and amplify templates containing uracil and inosine bases. This is useful for amplifying bisulfite-converted, enzymatically-deaminated, or damaged DNA, preventing carryover contamination in PCR (when used with dUTP and UDG), and in USER cloning methods. Learn more about this product.
- Highest fidelity amplification (>280X higher than Taq)
- Ultra-low error rates
- Superior performance for a broad range of amplicons (from high AT to high GC)
- Hot start and master mix formats available
The Q5 buffer system is designed to provide superior performance with minimal optimization across a broad range of amplicons, regardless of GC content. For routine or complex amplicons up to ~65% GC content, Q5 Reaction Buffer (NEB #B9027) provides reliable and robust amplification. For amplicons with high GC content (>65% GC), addition of the Q5 High GC Enhancer ensures continued maximum performance. Q5 and Q5 Hot Start DNA Polymerases are available as standalone enzymes, or in a master mix format for added convenience. Master mix formulations include dNTPs, Mg++ and all necessary buffer components.
In contrast to chemically modified or antibody-based hot start polymerases, NEB's Q5 Hot Start (NEB #M0493) utilizes a unique synthetic aptamer. This molecule binds to the polymerase through non-covalent interactions, blocking activity during the reaction setup. The polymerase is activated during normal cycling conditions, allowing reactions to be set up at room temperature. Q5 Hot Start does not require a separate high temperature activation step, shortening reaction times and increasing ease-of-use. Q5 Hot Start Polymerase is an ideal choice for high specificity amplification and provides robust amplification of a wide variety of amplicons, regardless of GC content.
Q5® is a registered trademark of New England Biolabs, Inc.
LabChip® is a registered trademark of Caliper Life Sciences, part of Perkin Elmer, Inc.
- PCR Using NEBNext® High-Fidelity 2X PCR Master Mix (M0541)
- PCR Using Q5® Hot Start High-Fidelity DNA Polymerase (M0493)
- Protocol for Q5® Hot Start High-Fidelity 2X Master Mix
- Protocol for Q5® High-Fidelity 2X Master Mix
- PCR Optimization (E0555)
- Protocol for a Routine PCR (E0555)
- PCR Using Q5® High-Fidelity DNA Polymerase (M0491)
- Protocol for a PCR reaction using NEBNext® Q5® Hot Start HiFi PCR Master Mix (M0543)
- Protocol for a PCR reaction using NEBNext® Ultra™ II Q5® Master Mix (M0544)
Anatomy of a Polymerase - How Function and Structure are Related
Read about the relationship between Polymerase structure and function when copying DNA.
Understanding Variability in DNA Amplification Reactions
- PCR Brochure
- DNA Polymerase Selection Chart
- PCR Troubleshooting Guide
- Guidelines for PCR Optimization with Thermophilic DNA Polymerases
- Amin Zargar, David N Quan, Milad Emamian, Chen Yu Tsao, Hsuan-Chen Wu, Chelsea R Virgile, William E Bentley 2015. Rational design of 'controller cells' to manipulate protein and phenotype expression. Metab Eng. , PubMedID: 25908186, DOI: 10.1016/j.ymben.2015.04.001
- Yuan Xue, Jossef Osborn, Anand Panchal, Jay L Mellies 2015. The RpoE Stress Response Pathway Mediates Reduction of the Virulence of Enteropathogenic Escherichia coli by Zinc. Appl Environ Microbiol. 81, PubMedID: 25819956, DOI: 10.1128/AEM.00507-15
- Longhai Dai, Can Liu, Yueming Zhu, Jiangsheng Zhang, Yan Men, Zeng Yan, Yuanxia Sun 2015. Functional Characterization of Cucurbitadienol Synthase and Triterpene Glycosyltransferase Involved in Biosynthesis of Mogrosides from Siraitia grosvenorii. Plant Cell Physiol. , PubMedID: 25759326, DOI: 10.1093/pcp/pcv043
- Jun Wu, Daiji Okamura, Mo Li, Keiichiro Suzuki, Chongyuan Luo, Li Ma, Yupeng He, Zhongwei Li, Chris Benner, Isao Tamura, Marie N Krause, Joseph R Nery, Tingting Du, Zhuzhu Zhang, Tomoaki Hishida, Yuta Takahashi, Emi Aizawa, Na Young Kim, Jeronimo Lajara, Pedro Guillen, Josep M Campistol, Concepcion Rodriguez Esteban, Pablo J Ross, Alan Saghatelian, Bing Ren, Joseph R Ecker, Juan Carlos Izpisua Belmonte 2015. An alternative pluripotent state confers interspecies chimaeric competency. Nature. , PubMedID: 25945737, DOI: 10.1038/nature14413
- Harish Nag Kankipati, Marta Rubio-Texeira, Dries Castermans, George Diallinas, Johan M Thevelein 2015. Sul1 and Sul2 Sulfate Transceptors Signal to Protein Kinase A upon Exit of Sulfur Starvation. J Biol Chem. 290, PubMedID: 25724649, DOI: 10.1074/jbc.M114.629022
- Yonghe Zhang, Huiming Huang, Shanshan Xu, Bo Wang, Jianhua Ju, Huarong Tan, Wenli Li 2015. Activation and enhancement of Fredericamycin A production in deepsea-derived Streptomyces somaliensis SCSIO ZH66 by using ribosome engineering and response surface methodology. Microb Cell Fact. 14, PubMedID: 25927229, DOI: 10.1186/s12934-015-0244-2
- Yafeng Li, Delu Song, Ying Song, Liangliang Zhao, Natalie Wolkow, John W Tobias, Wenchao Song, Joshua L Dunaief 2015. Iron-induced Local Complement Component 3 (C3) Up-regulation via Non-canonical Transforming Growth Factor (TGF)-β Signaling in the Retinal Pigment Epithelium. J Biol Chem. 290, PubMedID: 25802332, DOI: 10.1074/jbc.M115.645903
- Christine Henke, Pamela L Strissel, Maria-Theresa Schubert, Megan Mitchell, Claus C Stolt, Florian Faschingbauer, Matthias W Beckmann, Reiner Strick 2015. Selective expression of sense and antisense transcripts of the sushi-ichi-related retrotransposon - derived family during mouse placentogenesis. Retrovirology. 12, PubMedID: 25888968, DOI: 10.1186/s12977-015-0138-8
- Binyamin D Berkovits, Christine Mayr 2015. Alternative 3' UTRs act as scaffolds to regulate membrane protein localization. Nature. , PubMedID: 25896326, DOI: 10.1038/nature14321
- Martin Kostovcik, Craig C Bateman, Miroslav Kolarik, Lukasz L Stelinski, Bjarte H Jordal, Jiri Hulcr 2014. The ambrosia symbiosis is specific in some species and promiscuous in others: evidence from community pyrosequencing. ISME J. , PubMedID: 25083930, DOI: 10.1038/ismej.2014.115
- Bert De Rybel, Milad Adibi, Alice S. Breda, Jos R. Wendrich, Margot E. Smit, Ondej Novk, Nobutoshi Yamaguchi, Saiko Yoshida, Gert Van Isterdael, Joakim Palovaara, Bart Nijsse, Mark V. Boekschoten, Guido Hooiveld, Tom Beeckman, Doris Wagner, Karin Ljung, Christian Fleck, Dolf Weijers 2014. Integration of growth and patterning during vascular tissue formation in Arabidopsis Science. 345, PubMedID: 25104393, DOI: 10.1126/science.1255215
- Vidhyadhar Nandana, Sushant Singh, Abhay Narayan Singh, Vikash Kumar Dubey 2014. Procerain B, a cysteine protease from Calotropis procera, requires N-terminus pro-region for activity: cDNA cloning and expression with pro-sequence. Protein Expr Purif. 103C, PubMedID: 25173974, DOI: 10.1016/j.pep.2014.08.003
- Xin Duan, Arjun Krishnaswamy, Irina De la Huerta, Joshua R Sanes 2014. Type II Cadherins Guide Assembly of a Direction-Selective Retinal Circuit. Cell. 158, PubMedID: 25126785, DOI: 10.1016/j.cell.2014.06.047
- Connelly CM1, Porter LR2, TerMaat JR 2014. PCR amplification of a triple-repeat genetic target directly from whole blood in 15 minutes as a proof-of-principle PCR study for direct sample analysis for a clinically relevant target BMC Med Genet. 15, PubMedID: 25495904, DOI: 10.1186/s12881-014-0130-5
- Vladimir Potapov, Jennifer L. Ong. 2017. Examining Sources of Error in PCR by Single-Molecule Sequencing. PLOS One. , PubMedID: 28683110, DOI:
- Fidelity – the highest fidelity amplification available (>100X higher than Taq)
- Robustness – high specificity and yield with minimal optimization
- Coverage – superior performance for a broad range of amplicons (from high AT to high GC)
- Speed – short extension times
- Amplicon length – robust amplifications up to 20 kb for simple templates, and 10 kb for complex
Template/product specificity: Is RNA or DNA involved? Is the 3´ terminus at a gap, nick or at the end of the template?
Removal of existing nucleotides: Will the nucleotide(s) be removed from the existing polynucleotide chain as part of the protocol? If so, will they be removed from the 5´ or the 3´ end?
Thermal stability: Does the polymerase need to survive incubation at high temperature or is heat inactivation desirable?
Fidelity: Will subsequent sequence analysis or expression depend on the fidelity of the synthesized products?
This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
Here are some quick tips for getting the most out of NEB's Q5® High-Fidelity DNA Polymerase.
Not sure why Q5® is your best choice for high-fidelity amplification of GC-rich targets? NEB's scientists will show you why we call Q5 an "ultra-high fidelity polymerase".
Looking for tips on dealing with GC-bias in DNA amplification? NEB scientists have the expertise you need!
Make sure you're using the optimal polymerase for your DNA amplifications. Get tips on choosing the right DNA Polymerase for your application.