Product Class: Other

Q5® High-Fidelity DNA Polymerase
icon_cloned icon_recomb icon_hifi 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. 

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)
  • 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
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Product Information

Description

 

Q5® High-Fidelity DNA Polymerase is a 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. Q5 High-Fidelity DNA Polymerase is supplied with an optimized buffer system that allows robust amplification regardless of GC content. The 5X Q5 Reaction Buffer contains 2 mM Mg++ at final (1X) reaction concentrations and is recommended for most routine applications. For GC-rich targets (≥ 65% GC), amplification can be improved by the addition of the 5X Q5 High GC Enhancer. 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.

M0493 
Amplification of a variety of human genomic amplicons from low to high GC content using Q5 High-Fidelity DNA Polymerase. 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.

Advantages and Features

Application Features

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

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 74°C.

Storage Temperature

-20°C

Unit Assay Conditions

25 mM TAPS-HCl (pH 9.3 @ 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM β-mercaptoethanol, 200 μM dNTPs including [3H]-dTTP and 15 nM primed M13 DNA.

Protocols, Manuals & Usage

Protocols

  1. PCR Using Q5® High-Fidelity DNA Polymerase (M0491)

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® High-Fidelity DNA Polymerase?
  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 DNA Polymerase?
  6. My template is GC rich or supercoiled. How can I optimize my product yield using Q5® High-Fidelity DNA Polymerase?
  7. Do I need to modify my annealing temperature when using the Q5® High GC Enhancer?
  8. When should I add the High GC Enhancer?
  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. There is a precipitate in the bottom of the buffer tube. Is this normal?
  11. What length of product can be made by Q5® High-Fidelity DNA Polymerase?
  12. I am having trouble amplifying a template that is longer than 5kb. How can I optimize my product yield using Q5® High-Fidelity DNA Polymerase?
  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?
  17. Do other polymerases work in Q5® Reaction Buffer?
  18. I am competing in the iGEM competition. Do you have any products that I should consider from NEB?
  19. What is the difference between Q5® and Q5U™ Hot Start High-Fidelity DNA Polymerase?

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



Q5® is a registered trademark of New England Biolabs, Inc.