• My NEB
  • Print
  • PDF
  • OneTaq® 2X Master Mix with Standard Buffer



    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


    • 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


    Buffer 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


    Unit Assay Conditions

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

    Quality Control

    Quality Control Assays

    • PCR Amplification (DNA Polymerase):

      The polymerase master mix is tested in a polymerase chain reaction (PCR) using a control template and specific primers, resulting in the expected product.


    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.


    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

    Supporting Documents

    Material Safety Datasheets



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

    Selection Tools

    Usage Guidelines & Tips

    Troubleshooting Guides

    Interactive Tools

    Application Notes

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