Q5® Hot Start High-Fidelity 2X Master MixProduct information
Q5 Hot Start High-Fidelity 2X Master Mix
|100 rxns (2x1.25ml) ( 2 X )||-||Unavailable in your region|
Q5 Hot Start High-Fidelity 2X Master Mix
|500 rxns (1x12.5ml) ( 2 X )||-||Unavailable in your region|
Q5 Hot Start High-Fidelity 2X Master Mix
|500 rxns (10x1.25ml) ( 2 X )||-||Unavailable in your region|
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
Featured VideosView Video Library
Important Tips for Q5 Polymerase
Why Choose Q5 High-fidelity Polymerase?
Behind the paper: Examining Sources of Error in PCR by Single-Molecule Sequencing
Why is Tm Important in Primer Design?
Tips for Amplifying Large Amplicons
How to Amplify GC-rich DNA
5 Tips for Setting Up Your PCR
Overview of PCR
Quick Tips - What do I do if my master mix has a precipitate?
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.
Product SourceAn E. coli strain that carries the Q5 High-Fidelity DNA Polymerase gene.
Advantages and Features
- High-specificity PCR
- High-fidelity PCR
- Long or Difficult Amplification
- High-throughput PCR
Properties & Usage
- 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.
- Product specifications for individual components in Q5 Hot Start High-Fidelity 2X Master Mix are available separately.
- 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,
- 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,
- 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
Usage & Guidelines
Tools & Resources
FAQs & Troubleshooting
- What are the advantages to using Q5® Hot Start High-Fidelity 2X Master Mix?
- What is the fidelity of Q5® High-Fidelity DNA Polymerase?
- How should I determine an appropriate annealing temperature for my reaction?
- What should my primer concentration be when using Q5® High-Fidelity DNA Polymerase products?
- How should I set up a PCR reaction using Q5® Hot Start 2X Master Mix?
- My template is GC rich or supercoiled. How can I optimize my product yield using Q5® High-Fidelity Master Mixes?
- There is a precipitate in the bottom of the Master Mix tube? Is this normal?
- How do I activate Q5® Hot Start High-Fidelity DNA Polymerase?
- 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?
- What length of product can be made by Q5® High-Fidelity DNA Polymerase?
- I have a tube of the Q5® High GC Enhancer from another product formulation - can I add it to the Q5 Master Mix?
- What are the stability and storage requirements of the Q5® Master Mixes?
- Does Q5® High-Fidelity DNA Polymerase exhibit a strand displacement activity?
- Where can I find help troubleshooting my PCR?
- I'd like to clone a fragment amplified with Q5® High-Fidelity DNA Polymerase. Do I have to blunt-end clone?
- Will Q5® High-Fidelity DNA Polymerase incorporate dUTPs?
Citations & Technical Literature
Quality, Safety & Legal
Quality Assurance StatementQuality 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.
SpecificationsThe 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 AnalysisThe 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 DataSheetsThe following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
Q5® Hot Start High-Fidelity 2X Master Mix
Legal and DisclaimersThis 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.
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.
TrademarksNEW 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.
The supporting documents available for this product can be downloaded below.