Home PCR, qPCR & Amplification Technologies Q5 Quick-Load™ High-Fidelity 2X Master Mix

Q5 Quick-Load™ High-Fidelity 2X Master Mix

  • High fidelity amplification in a convenient format
  • Master mix includes Q5® High-Fidelity DNA Polymerase, dNTPs, and Mg++ in a broad-use buffer
  • Superior performance for a wide range of amplicons (from high AT to high GC)
  • Includes inert tracking dyes for easy and direct loading of PCR products onto gels
  • Compatible with a universal annealing temperature protocol at 60°C. Learn more in our application note.
  • Also available: Q5 standalone polymerase (NEB #M0491, NEB #M0493) and colorless master mix formats (NEB #M0492, NEB #M0494)
 
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Catalog # Cat # Concentration Conc Size List Price List Price Your Price Quantity
M0578L 2 X 500 reactions $868.00 $868.00
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  • [Duck photo] Learn more about the Q5® DNA Polymerase family of products


    The Q5 Quick-Load High-Fidelity 2X Master Mix offers high-fidelity performance in a convenient master mix format. The master mix features Q5, a high-fidelity, thermostable DNA polymerase with 3´to 5´ exonuclease activity fused to a processivity-enhancing Sso7d domain. With an error rate ~280-fold lower than that of Taq DNA Polymerase, Q5 offers highly accurate amplification and can be used to amplify difficult or long targets (up to 20 kb for simple templates). The master mix includes polymerase, dNTPs, Mg++, and two commonly used tracking dyes for easy and direct loading of PCR products onto gels. On a 1% agarose gel in 1X TBE, Xylene Cyanol FF migrates at ~3-5 kb and Tartrazine migrates at ~50 bp. The broad-use buffer requires only the addition of primers and DNA template for robust amplification, regardless of GC content.

    To determine the optimal annealing temperature for a given set of primers, use of the NEB Tm Calculator is highly recommended.


    Figure 1: Q5 formats support robust amplification of a broad range of targets

    Coverage diagram of Q5 High-Fidelity DNA Polymerases, with options for flexibility or convenience
    For flexibility, standalone Q5 polymerase products and the included High GC Enhancer can support amplification of targets with differing amounts of GC content. For convenience, and to amplify a broad range of target GC content, Q5 master mixes are recommended.

    Figure 2: Migration of tracking dyes contained in Quick-Load® PCR reagents during electrophoresis

    Green visible tracking dye with Quick-Load® separates to blue ~ 3-5 kb and yellow ~ 50 bp
    Quick-Load PCR reagents enable direct loading of PCR products onto agarose DNA gels for analysis. Quick-Load amplification products deliver the same performance as their non-Quick-Load equivalents, while incorporating two PCR-inert tracking dyes for enhanced visualization. During gel electrophoresis, the blue Xylene Cyanol FF dye co-migrates with DNA fragments of approximately 3-5 kb in a 1% agarose gel, while the yellow Tartrazine dye migrates at approximately 50 bp for clear monitoring of both large and small DNA fragments. Additionally, the combination of the blue and yellow dyes results in a green reagent that enables visual tracking of reaction set up.

    Figure 3: Q5 Quick-Load™ products can amplify long targets with high specificity and a simpler setup
    Gel depicting high performance amplification of long amplicons with Q5 Quick-Load High-Fidelity DNA Polymerase
    Triplicate PCR reactions were performed using two 5 kb human targets with 10 ng of human genomic DNA, utilizing Q5® High-Fidelity 2X Master Mix and Q5 Quick-Load High-Fidelity 2X Master Mix according to the manufacturer's recommended protocols. Both Q5 Quick-Load High-Fidelity 2X Master Mix and Q5 High-Fidelity 2X Master Mix demonstrated robust and highly specific amplification of long amplicons. L = Quick-Load® Purple 1 kb DNA Ladder (NEB #N0552).

    Figure 4: Q5 Quick-Load™ products exhibit equally high performance as Q5® products when amplifying AT-rich targets

    Gel depicting amplification of AT-rich targets with Q5 Quick-Load High-Fidelity DNA Polymerase
    Triplicate PCR reactions were performed using 4 human targets with a high AT content. The amplification was performed with 10 ng of human genomic DNA, utilizing Q5 High-Fidelity 2X Master Mix and Q5 Quick-Load High-Fidelity 2X Master Mix according to the manufacturer's recommended protocols. Both Q5 High-Fidelity 2X Master Mix and Q5 Quick-Load High-Fidelity 2X Master Mix consistently demonstrated robust and highly specific amplification of AT-rich amplicons. L = Quick-Load® Purple 1 kb DNA Ladder (NEB #N0552).

    Figure 5: Equally high performance on a broad range of amplicons with a convenient setup

    Gel depicting amplification of targets of medium lengths and different GC content with Q5 Quick-Load High-Fidelity DNA Polymerase
    Triplicate PCR reactions were performed using 4 human targets with average length and GC content. The amplification was performed with 10 ng of human genomic DNA, utilizing Q5® High-Fidelity 2X Master Mix and Q5 Quick-Load™ High-Fidelity 2X Master Mix according to the manufacturer's recommended protocols. Both Q5 High-Fidelity 2X Master Mix and Q5 Quick-Load High-Fidelity 2X Master Mix consistently demonstrated robust and highly specific amplification across a range of GC content. L = Quick-Load® Purple 1 kb DNA Ladder (NEB #N0552).

    [Icon] Use our Tm Calculator to determine your primers' optimal annealing temperature.

    This product is related to the following categories:
    Q5® High-Fidelity DNA Polymerases,
    Master Mixes,
    PCR, qPCR & Amplification Technologies
    This product can be used in the following applications:
    High-Fidelity PCR,
    DNA Amplification, PCR & qPCR,
    PCR

    • Reagents Supplied

      The following reagents are supplied with this product:

      NEB # Component Name Component # Stored at (°C) Amount Concentration

    • Properties & Usage

      Materials Required but not Supplied

      • Thermocycler
      • PCR tubes

    • Related Products

      Materials Sold Separately

  • Protocols, Manuals & Usage

  • Quality, Safety & Legal

    • Quality Control Assays

      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 & Change Notifications

      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 Data Sheets

      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.

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