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Measuring DNA cleavage levels has been a long-standing method in the field of topoisomerases. Due to the formation of a reversible covalent enzyme:DNA linkage, topoisomerase II activity can be monitored under varying conditions by means of measuring single- and double-stranded DNA breaks. This chapter will provide a method for measuring plasmid DNA cleavage levels generated by eukaryotic topoisomerase II with a specific focus on human type IIA topoisomerases. This method can be utilized to perform reactions as single timepoint, time course, and drug titrations.

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Plasmid DNA Cleavage Assay with Eukaryotic Topoisomerase II

  • 实验方案

  • 发表日期：

    2025年

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  • Allison J. Thomas,
  • Brooke D. Latham,
  • Addison K. O’Brian,
  • Mattalyn R. Hardin,
  • Joseph Deweese

摘要

Measuring DNA cleavage levels has been a long-standing method in the field of topoisomerases. Due to the formation of a reversible covalent enzyme:DNA linkage, topoisomerase II activity can be monitored under varying conditions by means of measuring single- and double-stranded DNA breaks. This chapter will provide a method for measuring plasmid DNA cleavage levels generated by eukaryotic topoisomerase II with a specific focus on human type IIA topoisomerases. This method can be utilized to perform reactions as single timepoint, time course, and drug titrations.