Q5® High-Fidelity DNA Polymerase sets a new standard for both fidelity and robust performance. With the highest fidelity amplification available (~280 times higher than Taq), Q5 results in ultra-low error rates. Q5 is composed of a novel polymerase that is fused to the processivity-enhancing Sso7d DNA binding domain, improving speed, fidelity and reliability of performance.
Working with uracil-containing DNA templates or using dUTP? Try Q5U® Hot Start High-Fidelity DNA Polymerase (NEB #M0515).
Highest fidelity amplification (~280X higher than Taq)
Master mix includes Q5 Hot Start High-Fidelity DNA Polymerase, dNTPs, and Mg++ in a broad-use buffer
Superior performance for a broad range of amplicons (from high AT to high GC)
Hot start aptamer offers increased reaction specificity with the benefit of room temperature setup
Bulk format available: M0494X = 1 x 12.5 ml bottle
Compatible with a universal annealing temperature protocol at 60°C. Learn more in our application note
The Q5® Hot Start High-Fidelity 2X
Master Mix features a high-fidelity, thermostable,
hot start DNA polymerase with 3´→ 5´ exonuclease
activity, fused to a processivity-enhancing Sso7d
domain to support robust DNA amplification. The
addition of an aptamer-based inhibitor allows
room temperature reaction setup. With an error rate ~280-fold lower than that of Taq DNA Polymerase, Q5 Hot Start High-Fidelity
2X Master Mix is ideal for cloning and can be used
for long or difficult amplicons. The convenient
master mix formulation is supplied at a 2X
concentration. The mix contains dNTPs, Mg++ and
a proprietary broad-use buffer requiring only the
addition of primers and DNA template for robust
amplification regardless of GC content. When used
at the recommended 1X final concentration, the
Q5 Hot Start High-Fidelity Master Mix contains
2 mM Mg++. Q5 Hot Start High-Fidelity 2X Master
Mix is unlike typical, lower fidelity PCR enzymes. To determine the optimal annealing temperatures for a given set of primers, use of the NEB Tm Calculator is highly recommended.
Figure 1: Q5 formats support robust amplification of a broad range of targets
For flexibility, standalone Q5 polymerase products and the included High GC Enhancer can support amplification of targets with differing amounts of GC content. For convenience, and to amplify a broad range of target GC content, Q5 master mixes are recommended. Figure 2: Amplification of a variety of human genomic amplicons from low to high GC content using Q5 High-Fidelity 2X Master Mix
All reactions were conducted using 30 cycles of amplification and visualized by microfluidic LabChip® analysis.
Product Source
An E. coli strain that carries the Q5 High-Fidelity DNA Polymerase gene.
This product is related to the following categories:
A precipitate (most noticeable after the first 1–2 freeze/thaw cycles) is not uncommon. To ensure optimal performance, the master mix should be thawed and resuspended prior to use. Stability testing using up to 15 freeze/thaw cycles has shown no negative effect on master mix performance.
Product specifications for individual components in Q5 Hot Start High-Fidelity 2X Master Mix are available separately.
References
Anastassia Voronova Erin Coyne, Ashraf Al Madhoun, Joel V. Fair, Neven Bosiljcic, Catherine St-Louis, Grace Li, Sherry Thurig, Valerie A. Wallace, Nadine Wiper-Bergeron, and Ilona S. Skerjanc. (2013). Hedgehog Signaling Regulates MyoD Expression and Activity. J Biol Chem. 288(6), 4389–4404. PubMedID: PMC3567689
Lieve Naesens, Luke Guddat, Dianne Keough, André B.P. van Kuilenburg, Judith Meijer, Johan Vande Voorde and Jan Balzarini (2013). ROLE OF HUMAN HYPOXANTHINE GUANINE PHOSPHORIBOSYLTRANSFERASE IN ACTIVATION OF THE ANTIVIRAL AGENT T-705 (FAVIPIRAVIR). Molecular Pharmacology Fast Forward. 87247,
Hicham Bouabe, Klaus Okkenhaug A Protocol for Construction of Gene Targeting Vectors and Generation of Homologous Recombinant Embryonic Stem Cells. Methods in Molecular Biology .
Thuronyi, B.W., Koblan, L.W., Levy, J.M. et al (2019) Continuous evolution of base editors with expanded target compatibility and improved activity Nat Biotechnol; 37, 1070-1079. PubMedID: 31332326, DOI: 10.1038/s41587-019-0193-0
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications & Change Notifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.
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.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
Licenses
This product is covered by one or more patents.
This product is licensed for research and commercial use from Bio-Rad Laboratories, Inc., under U.S. Pat. Nos. 6,627,424, 7,541,170, 7,670,808, 7,666,645, and corresponding patents in other countries. No rights are granted for use of the product for Digital PCR or real-time PCR applications, with the exception of quantification in Next Generation Sequencing workflows.
For additional information or to inquire about commercial use, please contact busdev@neb.com.
Trademarks
NEW ENGLAND BIOLABS® and Q5® are registered trademarks of New England Biolabs, Inc.
LABCHIP® is a registered trademark of Caliper Life Sciences, Inc., part of PerkinElmer, Inc.
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