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  • OneTaq® 2X Master Mix with Standard Buffer

    Description

    OneTaq® 2X Master Mix with Standard Buffer is an optimized blend of Taq and Deep VentR™ DNA Polymerases ideally suited to routine PCR applications from a variety of templates including pure DNA solutions, bacterial colonies, and cDNA products. The 3´→5´ exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA Polymerase (1). The convenient master mix formulation contains dNTPs, MgCl2, buffer components and stabilizers, requiring only the addition of primers and DNA template for robust amplification.

    Amplification of a selection of sequences with varying GC content from human and C. elegans genomic DNA using OneTaq 2X Master Mix with Standard Buffer. Amplicon sizes are indicated next to gel, and GC content is indicated above gel. Marker M is the 1 kb DNA Ladder.
      

    Product Source

    An E. coli strain that carries the Taq DNA Polymerase gene from Thermus aquaticus YT-1 and an E. coli strain that carries the Deep VentR DNA Polymerase gene from Pyrococcus species GB-D.

    Advantages and Features

    Applications

    • GC-rich PCR 
    • High Sensitivity PCR
    • High Throughput PCR 
    • Colony PCR 
    • Long PCR (up to ~6 kb genomic)
    • AT-rich PCR

    Properties and Usage

    Storage Temperature

    -20°C

    1X Master Mix Composition

    20 mM Tris-HCl
    22 mM KCl
    22 mM NH4Cl
    1.8 mM MgCl2
    5% Glycerol
    0.05% Tween® 20
    0.06% IGEPAL® CA-630
    0.2 mM dNTPs
    25 units/ml OneTaq® DNA Polymerase
    pH 8.9@25°C

    Heat Inactivation

    No

    Unit Assay Conditions

    1X ThermoPol® Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 200 µg/ml activated Calf Thymus DNA.

    Notes

    1. OneTaq 2X Master Mix with Standard Buffer is stable for fifteen freeze-thaw cycles when stored at -20°C.
    2. OneTaq 2X Master Mix with Standard Buffer is also stable for one month at 4°C, so for frequent use, an aliquot may be kept at 4°C.
    3. Product specifications for individual components in the OneTaq 2X Master Mix with Standard Buffer are available separately.

    References

    1. Barnes,W.M. (1994). Proc. Natl Acad. Sci. USA. 91, 2216-2220.
    2. Saiki,R.K. et al (1985). Science. 230, 1350-1354.
    3. Powell,L.M. et. al. (1987). Cell. 50, 831-840.
    4. Sun,y., Hegamyer, G. and Colburn,N. (1993). Biotechniques. 15, 372-374.
    5. Sarker, G., Kapelner, S. and Summer, S.S. (1990). Nuclear Acids Res.. 18, 7465.
    6. Magda Matoušková, Pavel Veselý, Petr Daniel, Giada Mattiuzzo, Ralph Hector, Linda Scobie, Yasuhiro Takeuchi and Jiří Hejnar (2013). The Role of DNA Methylation in Expression and Transmission of Procine Endogenous Retrovirus. Journal of Virology. JVI.03262-12

    FAQs

    1. How should I set up a PCR using the OneTaq® Master Mixes?
    2. Can I use my regular Taq-based cycling conditions for OneTaq® DNA Polymerase based products?
    3. Can I use OneTaq® DNA Polymerase in "hot start" PCR to help with specificity or primer dimer problems?
    4. What are the stability and storage requirements of the OneTaq® Master Mixes?
    5. Can OneTaq® DNA Polymerase be used in colony PCR?
    6. What type of DNA ends result from a primer extension reaction or a PCR using OneTaq® DNA Polymerase?
    7. How long a product can be made by OneTaq® DNA Polymerase?
    8. Can OneTaq® DNA Polymerase be used with uracil-containing primers or bisulfite-treated DNA?
    9. What is the fidelity of OneTaq® DNA Polymerase?
    10. Where can I find help troubleshooting my PCR?
    11. How should I determine an appropriate annealing temperature for my reaction?
    12. How is OneTaq DNA Polymerase different from LongAmp™ Taq DNA Polymerase?

    Tech Tips

    Use OneTaq 2X Master Mix with Standard Buffer to amplify routine targets with up to 65% GC content.

    Protocols

    1. Protocol for OneTaq® 2X Master Mix with Standard Buffer (M0482)

    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 Tools

    Usage Guidelines & Tips

    Troubleshooting Guides

    Application Notes

    Citations

    • Stöhr AC, López-Bueno A, Blahak S, Caeiro MF, Rosa GM, Alves de Matos AP, Martel A, Alejo A, Marschang RE (2015)Phylogeny and differentiation of reptilian and amphibian ranaviruses detected in europe PLoS One 10(2), e0118633. PubMedID: 25706285, DOI: 10.1371/journal.pone.0118633

    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.
    • 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.

    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.