Q5® High-Fidelity DNA Polymerases

Fidelity at its finest – for over 10 years.

Q5® High-Fidelity DNA Polymerase (NEB #M0491) sets a new standard for both fidelity and robust performance. With fidelity >280 times higher than Taq, Q5 results in ultra-low error rates. Q5 is a novel polymerase that is fused to the processivity-enhancing Sso7d DNA binding domain, improving speed, fidelity and reliability of performance.

Q5 is also available in a hot start format (NEB #M0493). In contrast to chemically modified or antibody-based hot start polymerases, Q5 Hot Start High-Fidelity DNA Polymerase utilizes engineered oligonucleotides known as aptamers. The aptamer binds to the polymerase through non-covalent interactions, blocking enzyme activity during the reaction setup, and is dissociated during normal cycling conditions. Reactions can be set up at room temperature and a separate high temperature activation step is not required, shortening reaction times.

Q5 master mixes (NEB #M0492, #M0494) contain dNTPs, Mg++ and a proprietary broad-use buffer requiring only the addition of primers and DNA template for robust amplification, regardless of GC content. 

Q5U® Hot Start High-Fidelity DNA Polymerase (NEB #M0515) 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.

The LunaScript® Multiplex One-Step RT-PCR Kit (NEB #E1555) combines Q5 with the thermostable Luna® WarmStart® Reverse Transcriptase for a streamlined one-step RT-PCR protocol with superior multiplexing and sensitivity down to 0.01 pg of human total RNA. Learn more about this product.

NEW: Q5 Blood Direct 2X Master Mix (NEB #M0500) can amplify a wide variety of targets direct from dried blood spots or up to 30% whole human blood, skipping DNA purification. Learn more about this product.

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The five features of Q5 High-Fidelity DNA Polymerase

  1. Extremely low error rates
    At ~280X higher than Taq, Q5 offers unparalleled fidelity for your most important samples.

  2. Robust amplification with minimal optimization
    High specificity and yield are absolute requirements for today's molecular biology techniques.

  3. Superior coverage for a broad range of amplicons, regardless of GC content
    While other DNA polymerases can have difficulty amplifying high-GC or high-AT amplicons, Q5 displays superior performance for a wide range of templates.

Robust Amplification with Q5 (A) and Q5 Hot Start (B) High-Fidelity DNA Polymerases

Amplification of a variety of human genomic amplicons from low to high GC content using either Q5 or Q5 Hot Start High-Fidelity DNA Polymerase. Reactions using Q5 Hot Start were set up at room temperature. All reactions were conducted using 30 cycles of amplification and visualized by microfluidic LabChip® analysis.

  1. Shorter PCR protocols
    Achieve precision without sacrificing speed. Q5's unique design incorporating the SSo7d processivity-enhancing domain enables shorter extension times, as low as 10 seconds per kb. Additionally, aptamer-based hot start requires no initial denaturation step and enables room temperature setup.

  2. Amplify templates up to 20 kb
    With Q5, you can reliably amplify simple templates up to 20 kb. Complex templates up to 10 kb can also be amplified with a high degree of confidence.


Master Mix and Stand-Alone Formats Provide Convenience and Flexibility


Q5® is a registered trademark of New England Biolabs, Inc.
LabChip® is a registered trademark of Caliper Life Sciences, part of Perkin Elmer, Inc.

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Protocols for Q5® High-Fidelity DNA Polymerases
Application Notes for Q5® High-Fidelity DNA Polymerases
    Publications related to Q5® High-Fidelity DNA Polymerases
    • Vladimir Potapov, Jennifer L. Ong. (2017) Examining Sources of Error in PCR by Single-Molecule Sequencing. PLOS One; PubMedID: 28683110
    • 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, 793-807. 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, 130. PubMedID: 25495904, DOI: 10.1186/s12881-014-0130-5
    • 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, 1255215. 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, 16-22. PubMedID: 25173974, DOI: 10.1016/j.pep.2014.08.003
    • 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, 10430-46. 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, 64. 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, 11918-34. 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, 9. 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
    • 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, 3766-74. PubMedID: 25819956, DOI: 10.1128/AEM.00507-15
Comparison of High-Fidelity Polymerases
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