Product Class: Other

Q5® Hot Start High-Fidelity 2X Master Mix
cloned at NEB recombinant No hifi hotstart pcr 3

Featured in the ARTIC SARS-CoV-2 sequencing workflow (1) and a component in the NEBNext® ARTIC SARS-CoV-2 kits.

Fidelity at its finest – for over 10 years

Product Introduction

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

Catalog # Size Concentration
M0494S 100.0 reactions 2 X
M0494L 500.0 reactions 2 X
M0494X 500.0 reactions 2 X

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Product Information

Description


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.
This product is related to the following categories:
Q5® High-Fidelity DNA Polymerases Products,
Master Mixes Products,
This product can be used in the following applications:
Site Directed Mutagenesis,
Site Directed Mutagenesis,
High-throughput cloning and automation solutions,
Hot Start PCR,
Long Range PCR,
Fast PCR,
High-Fidelity PCR,
Multiplex PCR,
Specialty PCR,
Routine PCR,
DNA Amplification, PCR & qPCR,
PCR,
Fast Cloning: Accelerate your cloning workflows with reagents from NEB

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Heat Inactivation

No

Application Features

  • High-specificity PCR
  • High-fidelity PCR
  • Cloning
  • Long or Difficult Amplification
  • High-throughput PCR

Product Notes

  1. 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.
  2. Product specifications for individual components in Q5 Hot Start High-Fidelity 2X Master Mix are available separately.

References

  1. 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.
  2. 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,
  3. Hicham Bouabe, Klaus Okkenhaug A Protocol for Construction of Gene Targeting Vectors and Generation of Homologous Recombinant Embryonic Stem Cells. Methods in Molecular Biology .

Protocols, Manuals & Usage

Protocols

  1. Protocol for Q5® Hot Start High-Fidelity 2X Master Mix

Usage & Guidelines

Application Notes

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. What are the advantages of using Q5® High-Fidelity DNA Polymerase?
  2. What are the differences between the numerous Q5® Polymerase products available?
  3. My results are not as expected. Where can I find troubleshooting help?
  4. What ends will my PCR products have?
  5. What does exonuclease activity mean for a DNA polymerase?
  6. Does Q5® High-Fidelity DNA Polymerase exhibit a strand displacement activity?
  7. What are the properties of this polymerase (fidelity, product ends, max amplicon, modified base incorporation, etc.)?
  8. How should I determine the appropriate annealing temperature for my reaction?
  9. What should the final primer concentration be in my PCR?
  10. When and how should I use the Q5® High GC Enhancer?
  11. There is a precipitate in the bottom of the buffer tube. Is this normal?
  12. What are Hot Start and WarmStart® polymerases and when would I use them?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

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 DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

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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