Product Class: Other

Q5® Hot Start High-Fidelity 2X Master Mix
icon_cloned icon_recomb icon_HI_no icon_hifi icon_hotstart icon_pcr tm+3

Product Introduction

Fidelity at its finest

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 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.

  • 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

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.
Product Categories:
Q5® High-Fidelity DNA Polymerases Products,
Master Mixes Products
Applications:
Site Directed Mutagenesis,
Site Directed Mutagenesis,
Hot Start PCR
,
Long Range PCR
,
Fast PCR
,
High-Fidelity PCR
,
DNA Amplification, PCR & qPCR
,
Multiplex PCR
,
Specialty PCR
,
Routine PCR
,
PCR
,
Fast Cloning: Accelerate your cloning workflows with reagents from NEB

Advantages and Features

Application Features

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

Properties & Usage

Storage Temperature

-20°C

Heat Inactivation

No

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. PubMedID: PMC3567689,
  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

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the Technical Support Form for appropriate document.)

Tools & Resources

Selection Charts

Web Tools

Faqs & Troubleshooting

FAQs

  1. What are the advantages to using Q5® Hot Start High-Fidelity 2X Master Mix?
  2. What is the fidelity of Q5® High-Fidelity DNA Polymerase?
  3. How should I determine an appropriate annealing temperature for my reaction?
  4. What should my primer concentration be when using Q5® High-Fidelity DNA Polymerase products?
  5. How should I set up a PCR reaction using Q5® Hot Start 2X Master Mix?
  6. My template is GC rich or supercoiled. How can I optimize my product yield using Q5® High-Fidelity Master Mixes?
  7. There is a precipitate in the bottom of the Master Mix tube? Is this normal?
  8. How do I activate Q5® Hot Start High-Fidelity DNA Polymerase?
  9. Are the DNA fragments produced by Q5® High-Fidelity DNA Polymerase blunt-ended or do they have the single-base 3´ overhang that Taq DNA Polymerase yields?
  10. What length of product can be made by Q5® High-Fidelity DNA Polymerase?
  11. I have a tube of the Q5® High GC Enhancer from another product formulation - can I add it to the Q5 Master Mix?
  12. What are the stability and storage requirements of the Q5® Master Mixes?
  13. Does Q5® High-Fidelity DNA Polymerase exhibit a strand displacement activity?
  14. Where can I find help troubleshooting my PCR?
  15. Will Q5® High-Fidelity DNA Polymerase incorporate dUTPs?
  16. I'd like to clone a fragment amplified with Q5® High-Fidelity DNA Polymerase. Do I have to blunt-end clone?

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

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].

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.

Licenses


This product is covered by one or more Patents.

This product is licensed 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 for use only in: (a) standard (not real-time) PCR in the research field only, but not real time PCR or digital PCR; (b) real-time PCR for use as a library preparation quantitation tool in Next Generation Sequencing workflows; (c) any in-vitro diagnostics applications, except for applications using real-time PCR or digital PCR; and (d) any non-PCR applications in DNA sequencing, isothermal amplification, and the production of synthetic DNA.

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.