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  • Uracil Glycosylase Inhibitor (UGI)

    Description

    The Uracil Glycosylase Inhibitor (UGI) of Bacillus subtilis bacteriophage PBS1 is a small protein (9.5 kDa) which inhibits E. coli uracil-DNA glycosylase (UDG) as well as UDG from other species. Inhibition of UDG occurs by reversible protein binding with a 1:1 UGD:UGI stoichiometry. UGI is capable of dissociating UDG-DNA complexes.


    Product Source

    An E. coli strain that carries the cloned UGI gene from Bacillus subtilis bacteriophage PBS1.

    Reagents Supplied

    The following reagents are supplied with this product:

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

    Advantages and Features

    Applications

    • Uracil glycosylase inhibitor (UGI) inhibits uracil-DNA glycosylase (UDG). Since UDG remains partially active following heat treatment at 95°C, UGI can be used to prevent subsequent degradation of product DNA.

    Properties and Usage

    Unit Definition

    One unit of UGI is defined as the amount of protein required to inhibit one unit of E. coli UDG in 1 hour at 37°C in a total reaction volume of 50 µl. One unit of UDG is the amount of enzyme which will catalyze the release of 60 pmol of uracil per minute from double stranded, uracil-containing DNA.

    Reaction Conditions

    1X UDG Reaction Buffer
    Incubate at 37°C

    1X UDG Reaction Buffer:
    20 mM Tris-HCl
    1 mM DTT
    1 mM EDTA
    pH 8 @ 25°C

    Storage Temperature

    -20°C

    Storage Conditions

    10 mM Tris-HCl
    50 mM KCl
    1 mM DTT
    0.1 mM EDTA
    200 μg/ml BSA
    50% Glycerol
    pH 7.4 @ 25°C

    Heat Inactivation

    No

    References

    1. Lindahl, T. et al. (1977). J. Biol. Chem.. 252, 3286-3294.
    2. Wang, Z., et al. (1991). Gene. 99, 31-37.
    3. Bennett, S. E. and Mosbaugh, D. W. (1992). J. Biol. Chem.. 267, 22512-22521.

    FAQs

    1. What is the nature of the Uracil Glycosylase Inhibitor?
    2. How does UGI inhibit UDG?
    3. Does UGI inhibit the activity of all UDGs?
    4. What is the molecular weight of UGI?
    5. Why is it necessary to use UGI in a reaction?

    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.

    Usage Guidelines & Tips

    Interactive Tools

    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.
    • Non-Specific DNase Activity (16 hour):
      The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.

    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.