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  • Bst DNA Polymerase, Full Length

    Description

    Bst DNA Polymerase, Full Length is the full length polymerase from Bacillus stearothermophilus. It has 5´ → 3´ polymerase and double-strand specific 5´ → 3´ exonuclease activity, but lacks 3´ → 5´ exonuclease activity (1).

    Product Source

    An E. coli strain that contains the gene from Bacillus stearothermophilus. (H. Kong)

    Reagents Supplied

    The following reagents are supplied with this product:

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

    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 65°C.

    Reaction Conditions

    1X ThermoPol® Reaction Buffer
    Incubate at 65°C

    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 Conditions

    10 mM Tris-HCl
    50 mM KCl
    1 mM DTT
    0.1 mM EDTA
    50% Glycerol
    0.1% Triton® X-100
    pH 7.5 @ 25°C

    Heat Inactivation

    80°C for 20 min

    5' - 3' Exonuclease

    Yes

    3' - 5' Exonuclease

    No

    Strand Displacement

    No

    Unit Assay Conditions

    50 mM KCl, 20 mM Tris-HCl (pH 8.8), 10 mM MgCl2, 30 nM M13mp18 SS DNA, 70 nM M13 sequencing primer (–47) 24-mer, 200 µM dATP, 200 µM dCTP, 200 µM dGTP, 100 µM dTTP including [3H]-dTTP and 100 µg/ml BSA.

    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.
    • Protein Purity (SDS-PAGE):
      The physical purity is assessed by comparing contaminating protein bands in a concentrated sample to the protein of interest band in a sample of known dilution. The purity is determined by SDS-PAGE.

    Notes

    1. Bst DNA Polymerase does not exhibit 3´→ 5´ exonuclease activity.
    2. Reaction temperatures above 70°C are not recommended.
    3. Cannot be used for thermal cycle sequencing or PCR.

    References

    1. Aliotta, J.M. et al. (1996). Genetic Analysis. 12, 185-195.
    2. Mead, D.A. et al. (1991). BioTechniques. 11, 76-87.

    Supporting Documents

    Material Safety Datasheets

    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.

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