Plasmid DNA Cleavage Assay with Eukaryotic Topoisomerase II.

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2025-01-01 DOI:10.1007/978-1-0716-4550-5_8

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

引用次数: 0

Abstract

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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真核生物拓扑异构酶ⅱ质粒DNA切割试验。

在拓扑异构酶领域，测量DNA切割水平一直是一种长期存在的方法。由于形成了一种可逆的共价酶：DNA链，拓扑异构酶II的活性可以通过测量单链和双链DNA断裂来监测在不同条件下。本章将提供一种测量真核生物拓扑异构酶II产生的质粒DNA切割水平的方法，并特别关注人类IIA型拓扑异构酶。该方法可用于单时间点、时间过程和药物滴定。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.