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Related Experiment Videos

General method for quantifying base adducts in specific mammalian genes.

D C Thomas1, A G Morton, V A Bohr

  • 1Department of Biochemistry, University of North Carolina School of Medicine, Chapel Hill 27599-7260.

Proceedings of the National Academy of Sciences of the United States of America
|June 1, 1988
PubMed
Summary
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A new method quantifies DNA adducts in mammalian genomes using ABC excinuclease. This technique measures DNA adduct formation and removal in specific genomic sequences.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • DNA adducts are critical markers of DNA damage and repair.
  • Quantifying DNA adducts in specific genomic regions is essential for understanding DNA repair mechanisms.

Purpose of the Study:

  • To develop a general method for measuring DNA adduct formation and removal in defined mammalian genomic sequences.

Main Methods:

  • Genomic DNA is digested and treated with Escherichia coli UvrABC excision nuclease (ABC excinuclease).
  • Alkaline gel electrophoresis and Southern hybridization are used to probe specific sequences.
  • ABC excinuclease incises adducted DNA, reducing fragment intensity proportionally to adduct number.

Main Results:

Related Experiment Videos

  • The method allows for the quantification of DNA adducts in specific genomic sequences.
  • The intensity reduction in Southern hybridization directly correlates with the number of DNA adducts.

    • Conclusions:

    • This method provides a versatile tool for studying DNA adducts across various genomic locations.
    • The wide substrate range of ABC excinuclease enables the quantification of diverse DNA adducts.