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Q5® High-Fidelity DNA Polymerases

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.

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Comparison of High-Fidelity Polymerases

1 We continue to investigate improved assays to characterize Q5’s very low error rate to ensure that we present the most accurate fidelity data possible (Potapov, V. and Ong, J.L. (2017) PLoS ONE. 12(1): e0169774).
2 PCR-based mutation screening in lacI (Agilent) or rpsL (Life).

Advantages

  • 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.

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.

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.

Amplification Performance Across a Wide Range of Genomic Targets

PCR was performed with a variety of amplicons, with GC content ranging from high AT to high GC, with Q5 and several other commercially available polymerases. All polymerases were cycled according to manufacturer's recommendations, including use of GC Buffers and enhancers when recommended. Yield and purity of reaction products were quantitated and represented, as shown in the figure key, by dot color and size. A large dark green dot represents the most successful performance. Q5 provides superior performance across the range of GC content.

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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FAQs for Q5® High-Fidelity DNA Polymerases

Protocols for Q5® High-Fidelity DNA Polymerases

Application Notes for Q5® High-Fidelity DNA Polymerases

    Publications related to Q5® High-Fidelity DNA Polymerases

  1. 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
  2. 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
  3. 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
  4. 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
  5. Binyamin D Berkovits, Christine Mayr 2015. Alternative 3' UTRs act as scaffolds to regulate membrane protein localization. Nature. , PubMedID: 25896326, DOI: 10.1038/nature14321
  6. 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
  7. 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
  8. 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
  9. 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
  10. 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
  11. 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
  12. 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
  13. 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
  14. 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

PCR Selection Tool

Choose from one of the largest selections of polymerases for PCR applications from the leader in enzyme technology and bring unparalleled confidence to your experiments.

Comparison of High-Fidelity Polymerases

Q5 DNA Polymerase Offers Superior Amplification for a Wide Range of Templates

*Regardless of GC content
Amplification of a variety of human genomic amplicons from low to high GC content demonstrates the broad performance of Q5 High-Fidelity DNA Polymerase. All reactions were conducted using 20 ng of input template and included 30 cycles of amplification. Results were visualized by microfluidic LabChip® analysis. Competitor polymerases were cycled according to manufacturer's recommendations. For the final three amplicons, GC Buffers or enhancers were used when supplied with the polymerase.

Five Quality Features of Q5

  1. Fidelity – the highest fidelity amplification available (>100X higher than Taq)
  2. Robustness – high specificity and yield with minimal optimization
  3. Coverage – superior performance for a broad range of amplicons (from high AT to high GC)
  4. Speed – short extension times
  5. Amplicon length – robust amplifications up to 20 kb for simple templates, and 10 kb for complex

DNA Polymerase Selection Chart

NEB offers a guidelines for choosing the correct DNA polymerase for your application by providing a list of specific properites.
Several factors govern which polymerase should be used in a given application, including: 

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?

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