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Q5® Hot Start High-Fidelity 2X Master Mix

Catalog # Concentration Size List Price Quantity Your Price
M0494L 2 X 500 reactions $1,227.00
$1,104.30
M0494S 2 X 100 reactions $309.00
$278.10
M0494X 2 X 500 reactions $1,227.00
$1,104.30
Catalog # Size List Price Your Price
M0494L 500 reactions $1,227.00
$1,104.30
M0494S 100 reactions $309.00
$278.10
M0494X 500 reactions $1,227.00
$1,104.30
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

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? Learn about 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)
  • Bulk format available: M0494X = 1 x 12.5 ml bottle

1. Josh Quick 2020. nCoV-2019 sequencing protocol v3 (LoCost). protocols.io https://www.protocols.io/view/ncov-2019-sequencing-protocol-v3-locost-bh42j8ye

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    Quick Tips - What do I do if my master mix has a precipitate?


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.

Q5-High Fidelity 2X Master Mix formats allow robust amplification of a broad range of targets with a single formulation. 

M0494
Amplification of a variety of human genomic amplicons from low to high GC content using Q5 Hot Start High-Fidelity 2X Master Mix. All reactions were set up at room temperature, 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.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Q5® Hot Start High-Fidelity 2X Master Mix M0494SVIAL -20 2 x 1.25 ml 2 X
  Q5® Hot Start High-Fidelity 2X Master Mix M0494SVIAL -20 10 x 1.25 ml 2 X
  Q5® Hot Start High-Fidelity 2X Master Mix M0494XVIAL -20 1 x 12.5 ml 2 X
Application Features
  • High-specificity PCR
  • High-fidelity PCR
  • Cloning
  • Long or Difficult Amplification
  • High-throughput PCR

Properties & Usage

Heat Inactivation

No

Companion Products
Notes
  • 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 .
Citations
Publications
  • 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.. DOI: 10.1038/s41587-019-0193-0
Quality Control Assay
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
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]
Certificate of Analysis
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]
Safety Data Sheets
Legal And Disclaimer

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.

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.

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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