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  • Therminator™ DNA Polymerase

    Did you know this product can be customized or purchased in larger volumes? Submit an inquiry to find out more.
    cloned at neb recombinant unique buffer heat inactivation no
    Catalog #SizeConcentrationPriceQtyAdd to Cart
    M0261S200 units2,000 units/ml$96.00Add to Cart
    M0261L1,000 units2,000 units/ml$384.00Add to Cart
      
    Categories:
    DNA Labeling

    Description

    Therminator DNA Polymerase is a 9°N™ DNA Polymerase variant with an enhanced ability to incorporate modified substrates such as dideoxynucleotides, ribonucleotides and acyclonucleotides (1,2).

    Highlights

    • Isolated from a recombinant source
    • Supplied with 10X Reaction Buffer
    • Enhanced incorporation of modified nucleotides
    • Single base incorporation of dideoxy and acyclonucleotides

    Product Source

    An E. coli strain that carries the 9°N (D141A / E143A / A485L) DNA Polymerase gene, a genetically engineered form of the native DNA polymerase from Thermococcus species 9°N-7.

    Reagents Supplied

    The following reagents are supplied with this product:

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

    Advantages and Features

    Applications

    • DNA sequencing by partial ribosubstitution (3)
    • DNA sequencing using dideoxy (4) or acyclo (5) chain terminators
    • SNP analysis with dideoxy or acyclo chain terminators (6)

    Properties and Usage

    Unit Definition

    One unit is defined as the amount of enzyme that will incorporate 10 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
    50% Glycerol
    pH 7.4 @ 25°C

    Heat Inactivation

    No

    Molecular Weight

    Theoretical: 90000 daltons

    5' - 3' Exonuclease

    No

    3' - 5' Exonuclease

    No

    Strand Displacement

    +

    Unit Assay Conditions

    1X ThermoPol Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 15 nM primed single-stranded M13mp18.

    References

    1. Gardner, A.F. and Jack, W.E. (1999). Nucl. Acids Res.. 27, 2545-2555.
    2. Gardner, A.F. and Jack, W.E. (2002). Nucl. Acids Res. 30, 605-613.
    3. Barnes, W.F. (1978). J. Mol. Biol.. 119, 83-99.
    4. Sanger, F., Nicklen, S. and Coulson, A.R. (1977). Proc. Natl. Acad. Sci. USA. 74, 5463-5467.
    5. Trainor, G.L. (1996). U.S. Patent No. 5,558,991.
    6. Haff, L.A. and Simirnov, I.P. (1997). Genome Methods. 7, 378-388.

    FAQs

    1. What is the specific activity of Therminator DNA Polymerase?
    2. Can Therminator DNA Polymerase be used in other buffers?
    3. When should Therminator DNA Polymerase be used?
    4. Can Therminator DNA Polymerase be substituted for ThermoSequenase?
    5. Can Therminator DNA Polymerase be used for sequencing?
    6. Why are some of the A's in my sequence weak when using Therminator DNA Polymerase?
    7. Does Therminator DNA Polymerase have 3'→5' proofreading exonuclease activity?

    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.
    • Exonuclease Activity (Radioactivity Release):
      The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.

    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.