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    Taq DNA Polymerase with ThermoPol® Buffer

    Description

      
    Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1,2,3) and a 5´ flap endonuclease activity (4,5).

    It is supplied with 10X ThermoPol Reaction Buffer, which contains a nonionic detergent to increase enzyme stability during longer incubations.

    Highlights

    • Isolated from a recombinant source
    • Supplied with 10X Reaction Buffer
    • Robust and reliable reactions
    • Tolerates a wide range of templates
    • Incorporates dUTP, dITP and fluorescently-labeled nucleotides
    • Exceptional value in terms of cost per unit

    Product Source

    An E. coli strain that carries the Taq DNA Polymerase gene from Thermus aquaticus YT-1

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    ThermoPol® Reaction Buffer-2010X

    Advantages and Features

    Applications

    • PCR
    • Primer extension
    • Colony PCR

    Properties and Usage

    Unit Definition

    One unit is defined as the amount of enzyme that will incorporate 15 nmol of dNTP into acid-insoluble material in 30 minutes at 75°C.

    Reaction Conditions

    1X ThermoPol® Reaction Buffer

    1X ThermoPol® Reaction Buffer:
    20 mM Tris-HCl
    10 mM (NH4)2SO4
    10 mM KCl
    2 mM MgSO4
    0.1% Triton® X-100
    pH 8.8 @ 25°C

    Storage Temperature

    -20°C

    Storage Conditions

    10 mM Tris-HCl
    100 mM KCl
    1 mM DTT
    0.1 mM EDTA
    0.5% Tween® 20
    0.5% IGEPAL® CA-630
    50% Glycerol
    pH 7.4 @ 25°C

    Heat Inactivation

    No

    Molecular Weight

    Theoretical: 94000 daltons

    5' - 3' Exonuclease

    Yes

    3' - 5' Exonuclease

    No

    Strand Displacement

    +1

    Resulting Ends

    • Single-base 3´ Overhangs

    Unit Assay Conditions

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

    Error Rate

    ~ 285x10-6bases

    Notes

    1. 5'→3' flap endonuclease activity destroys displaced strand.

    References

    1. Chien, A., Edgar, D.B. and Trela, J.M. (1976). J. Bact.. 127, 1550-1557.
    2. Kaledin, A.S., Sliusarenko, A.G. and Gorodetskii, S.I. (1980). Biokhimiya. 45, 644-651.
    3. Lawyer, F.C. et al. (1993). PCR Methods and Appl.. 2, 275-287.
    4. Longley, M.J., Bennett, S.E. and Mosbaugh D.W. (1990). Nucleic Acids Res.. 18, 7317-7322.
    5. Lyamichev, V., Brow, M.A. and Dahlberg, J.E. (1993). Science. 260, 778-783.
    6. Saiki R.K. et al. (1985). Science. 230, 1350-1354.
    7. Powell, L.M. et al. (1987). Cell. 50, 831-840.
    8. Sun, Y., Hegamyer, G. and Colburn, N. (1993). Biotechniques. 15, 372-374.
    9. Sarkar, G., Kapelner, S. and Sommer, S.S. (1990). Nucleic Acids Res.. 18, 7465.

    FAQs

    1. Can Taq DNA Polymerase be used in other buffers?
    2. Why is the product a smear when visualized on an agarose gel?
    3. Why is there no product when visualized on an agarose gel?
    4. How should I set up an amplification reaction using Taq DNA Polymerase?
    5. What type of DNA ends result from a primer extension reaction or a PCR using Taq DNA Polymerase?
    6. The product sequence doesn't completely match the expected sequence. How can this result be improved?
    7. When should Taq DNA Polymerase be used in a primer extension reaction or for PCR?
    8. Will the 5'→3' flap endonuclease activity of Taq DNA Polymerase degrade primers?
    9. Can Taq DNA Polymerase be used for nick translation?
    10. Does the presence of Ca2+ inhibit PCR?

    Tech Tips

    Did you know most Taq reactions amplify more efficiently and robustly when you use a 68°C extension temperature?

    Protocols

    1. PCR Protocol for Taq DNA Polymerase with ThermoPol® Buffer (M0267)
    2. A-Tailing with Taq Polymerase

    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

    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.
    • Endonuclease Activity (Nicking):
      The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
    • 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.
    • Single Stranded DNase Activity (FAM Labeled Oligo):
      The product is tested in a reaction containing a fluorescent internal labeled single stranded oligonucleotide. The percent degradation is determined by capillary electrophoresis.

    Material Safety Data Sheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

    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.