Q5® High-Fidelity 2X Master Mix

cloned at neb recombinant heat inactivation no hifi pcr tm+3
Catalog #SizeConcentrationPriceQtyAdd to Cart
M0492S100 reactions (50 μl vol)2X$172.00Add to Cart
M0492L500 reactions (50 μl vol)2X$688.00Add to Cart
  
Categories:
Master Mix Products,
Master Mix Products,
Q5® High-Fidelity DNA Polymerase Products
Applications:
DNA Amplification, PCR and qPCR,
Fast PCR,
Fast PCR

Description

  
The Q5® High-Fidelity 2X Master Mix offers robust, high-fidelity performance in a convenient master mix format. The Q5 High- Fidelity 2X Master Mix features 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 > 100-fold lower than that of Taq DNA Polymerase and 12-fold lower than that of Pyrococcus furiosus (Pfu) DNA Polymerase, Q5 High-Fidelity DNA Polymerase 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 High-Fidelity Master Mix contains 2 mM Mg++. 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.

Q5-High Fidelity 2X Master Mix formats allow robust amplification of a broad range of targets with a single formulation. 

M0492
Amplification of a variety of human genomic amplicons from low to high GC content using Q5 High-Fidelity 2X Master Mix. 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

Applications

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

Properties and Usage

Storage Temperature

-20°C

Heat Inactivation

No

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 the Q5 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 (2013). A Protocol for Construction of Gene Targeting Vectors and Generation of Homologous Recombinant Embryonic Stem Cells. Methods in Molecular Biology. 1064, 337-354.

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. How should I set up a PCR experiment using Q5® High-Fidelity 2X Master Mix?
  5. What should my primer concentration be when using Q5® High-Fidelity DNA Polymerase products?
  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. 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?
  9. What length of product can be made by Q5® High-Fidelity DNA Polymerase?
  10. I have a tube of the Q5® High GC Enhancer from another product formulation - can I add it to the Q5 Master Mix?
  11. What are the stability and storage requirements of the Q5® Master Mixes?
  12. Does Q5® High-Fidelity DNA Polymerase exhibit a strand displacement activity?
  13. Where can I find help troubleshooting my PCR?
  14. Will Q5® High-Fidelity DNA Polymerase incorporate dUTPs?
  15. I'd like to clone a fragment amplified with Q5® High-Fidelity DNA Polymerase. Do I have to blunt-end clone?

Protocols

  1. Protocol for Q5® High-Fidelity 2X Master Mix

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 info@neb.com or fill out the Technical Support Form for appropriate document.

Selection Charts

Feature Articles

Usage Guidelines & Tips

Troubleshooting Guides

Interactive Tools

Application Notes

Quality Control

Quality Control Assays

The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page. Further information regarding NEB product quality can be found here.
  • PCR Amplification (Master Mix):
    The polymerase master mix is tested in a polymerase chain reaction (PCR) using a control template and specific primers, resulting in the expected product.

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]

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

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 info@neb.com or fill out the Technical Support Form for appropriate document.