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  • Hemo KlenTaq®

    Description

    Hemo KlenTaq is a truncated version of Taq DNA Polymerase, lacking the first 280 amino acids (1). Hemo KlenTaq also contains mutations that make it resistant to inhibitors present in whole blood. It can amplify directly from whole blood from humans and mice. Hemo KlenTaq tolerates up to 20% whole blood in a 25 µl reaction (30% in 50 µl reaction).

    Amplification of human whole blood with Hemo KlenTaq. Percent blood present in sample and anticoagulant used are indicated in the legend. Ladder M is the 2-Log DNA Ladder (NEB #N3200).

    Product Source

    An E. coli strain that carries a mutant Taq DNA polymerase gene. The protein lacks the N-terminal 5´→ 3´ exonuclease domain and the gene has three internal point mutations.

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    Hemo KlenTaq® Reaction Buffer Pack-205X

    Advantages and Features

    Features

    • Versatility and robustness of Taq DNA Polymerase to amplify DNA from whole blood.

    Applications

    • Whole Blood PCR
    • Dried Blood Spot PCR
    • Primer extension

    Properties and Usage

    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

    5' - 3' Exonuclease

    No

    3' - 5' Exonuclease

    No

    Resulting Ends

    • Single-base 3´ Overhangs

    Unit Assay Conditions

    1X ThermoPol® Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 15 nM primed M13 DNA.

    References

    1. Kermekchiev, M.B., et al. (2009). Nucleic Acids Res. 37, e40.
    2. Sun, Y., Hegamyer, G. and Colburn, N. (1993). Biotechniques . 15, 372-374.

    Tech Tips

    For best results, slowly pipette your blood sample into the reaction last and let it settle to the bottom of the reaction. The reaction should not be mixed once the blood is added.

    Protocols

    1. PCR Protocol for Hemo KlenTaq®

    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.

    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 (Whole Blood DNA):
      The polymerase is tested in a polymerase chain reaction (PCR) using whole blood containing inhibitors and specific primers, resulting in the expected product.
    • Single Stranded DNA Binding (FAM labeled oligo):
      The product is tested for its ability to produce a mobility shift when incubated using standard reaction conditions with a single stranded FAM labeled oligo.

    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.