Quantification of Double-Strand Breaks in Mammalian Cells Using Pulsed-Field Gel Electrophoresis

Kelvin W Pond; Nathan A Ellis

doi:10.1007/978-1-4939-9500-4_4

Quantification of Double-Strand Breaks in Mammalian Cells Using Pulsed-Field Gel Electrophoresis

Methods Mol Biol. 2019:1999:75-85. doi: 10.1007/978-1-4939-9500-4_4.

Authors

Kelvin W Pond¹, Nathan A Ellis²

Affiliations

¹ Department of Cellular and Molecular Medicine, University of Arizona Cancer Center, University of Arizona, Tuscon, AZ, USA.
² Department of Cellular and Molecular Medicine, University of Arizona Cancer Center, University of Arizona, Tuscon, AZ, USA. naellis@email.arizona.edu.

PMID: 31127570
DOI: 10.1007/978-1-4939-9500-4_4

Abstract

The double-strand break (DSB) is the most cytotoxic type of DNA damage and measurement of DSBs in cells is essential to understand their induction and repair. Pulsed-field gel electrophoresis (PFGE) allows for quantitative measurement of DSBs in a cell population generated by DNA damaging agents. PFGE has the capacity to separate DNA molecules from several hundred base pairs to over six million base pairs. In the method described here, molecules from five hundred thousand to three million base pairs are consolidated into a single band on the gel that is readily analyzed.

Keywords: Agarose gel; DNA damage; Double-strand breaks; Intercalating agent; Pulsed-field gel electrophoresis; Radiation.

MeSH terms

Animals
Cells, Cultured
DNA / genetics
DNA / isolation & purification*
DNA Breaks, Double-Stranded / drug effects
DNA Breaks, Double-Stranded / radiation effects
DNA Repair
Electrophoresis, Gel, Pulsed-Field / methods*
Gamma Rays / adverse effects
Humans
Intercalating Agents

Substances

Intercalating Agents
DNA