Product Class: Kit

Q5® High-Fidelity PCR Kit

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

  

Q5® High-Fidelity DNA Polymerase is an ultra high-fidelity, thermostable DNA polymerase with 3´→ 5´ exonuclease activity, fused to a processivity-enhancing Sso7d domain to support robust DNA amplification. With an error rate ~280-fold lower than that of Taq DNA Polymerase, Q5 High-Fidelity DNA Polymerase is ideal for cloning and can be used for long or difficult amplicons. The Q5 High-Fidelity 2X Master Mix offers robust, high-fidelity performance in a convenient master mix format. 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 High-Fidelity Master Mix contains 2 mM MgCl2.

Q5 High-Fidelity DNA Polymerase 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.

This kit contains a sufficient supply of master mix and nuclease-free water to perform 50 polymerase chain reactions (NEB #E0555S) or 200 reactions (NEB #E0555L). For added convenience, gel loading dye, and a Quick-Load DNA LAdder are also included in this kit.

E0555

Kit Components

The following reagents are supplied with this product:

Store at (°C) Concentration
Nuclease-free Water -20
Gel Loading Dye, Purple (6X) 25 6 X
Q5® High-Fidelity 2X Master Mix -20 2 X
Quick-Load® Purple 1 kb Plus DNA Ladder 4
Product Categories:
Q5® High-Fidelity DNA Polymerases Products
Applications:
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

Properties & Usage

Product Notes

  1. Product specifications for individual components in the Q5 High-Fidelity PCR Kit 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. (2013). A Protocol for Construction of Gene Targeting Vectors and Generation of Homologous Recombinant Embryonic Stem Cells. Methods in Molecular Biology . 1064, 337-354.

Protocols, Manuals & Usage

Protocols

  1. Protocol for a Routine PCR (E0555)
  2. PCR Optimization (E0555)

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

Faqs & Troubleshooting

FAQs

  1. What are the advantages to using Q5® 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 experiment using Q5® High-Fidelity 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. I have a tube of the Q5® High GC Enhancer from another product formulation - can I add it to the Q5 Master Mix?
  8. There is a precipitate in the bottom of the Master Mix tube? Is this normal?
  9. What are the stability and storage requirements of the Q5® PCR Kit?
  10. 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?
  11. What length of product can be made by Q5® High-Fidelity DNA Polymerase?
  12. Does Q5® High-Fidelity DNA Polymerase exhibit a strand displacement activity?
  13. Will Q5® High-Fidelity DNA Polymerase incorporate dUTPs?
  14. How much of the DNA Ladder should I load on my gel?
  15. Where can I find additional help troubleshooting my PCR?

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.