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Q5® Hot Start High-Fidelity DNA Polymerase
cloned at NEB recombinant No hifi hotstart pcr 3

Fidelity at its finest – for over 10 years

This product is available in a glycerol-free format. Contact us for more information.

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)
  • Ultra-low error rates
  • Superior performance for a broad range of amplicons (from high AT to high GC)
  • Also available: Q5 standalone polymerase (NEB #M0491) and master mix formats (NEB #M0492, M0494)
Catalog # Size Concentration
M0493S 100.0 units 2000 units/ml
M0493L 500.0 units 2000 units/ml

Featured Videos

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    Behind the paper: Examining Sources of Error in PCR by Single-Molecule Sequencing

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Product Information

Description

 

Q5® Hot Start High-Fidelity DNA Polymerase is 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 DNA Polymerase is ideal for cloning and can be used for long or difficult amplicons. Q5 Hot Start 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 Hot Start 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 Hot Start High-Fidelity DNA Polymerase. 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,
This product can be used in the following applications:
Hot Start PCR,
Long Range PCR,
Fast PCR,
High-Fidelity PCR,
Genome Editing Applications,
Multiplex PCR,
Specialty PCR,
DNA Amplification, PCR & qPCR,
Routine PCR,
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

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.

Heat Inactivation

No

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.

Application Features

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

Protocols, Manuals & Usage

Protocols

  1. PCR Using Q5® Hot Start High-Fidelity DNA Polymerase (M0493)

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. What should the final primer concentration be in my PCR?
  9. When and how should I use the Q5® High GC Enhancer?
  10. Do other polymerases work in Q5® Reaction Buffer?
  11. There is a precipitate in the bottom of the Master Mix 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.
SYBR® is a registered trademark of Molecular Probes, Inc., now owned by Life Technologies, Inc.

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