Quantification of double-strand breaks in mammalian cells using pulsed-field gel electrophoresis

Kelvin W. Pond, Nathan Ellis

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages75-85
Number of pages11
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1999
ISSN (Print)1064-3745

Fingerprint

Pulsed Field Gel Electrophoresis
Base Pairing
DNA
DNA Damage
Gels
Population
1,2-di-(4-sulfamidophenyl)-4-butylpyrazolidine-3,5-dione

Keywords

  • Agarose gel
  • DNA damage
  • Double-strand breaks
  • Intercalating agent
  • Pulsed-field gel electrophoresis
  • Radiation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Pond, K. W., & Ellis, N. (2019). Quantification of double-strand breaks in mammalian cells using pulsed-field gel electrophoresis. In Methods in Molecular Biology (pp. 75-85). (Methods in Molecular Biology; Vol. 1999). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9500-4_4

Quantification of double-strand breaks in mammalian cells using pulsed-field gel electrophoresis. / Pond, Kelvin W.; Ellis, Nathan.

Methods in Molecular Biology. Humana Press Inc., 2019. p. 75-85 (Methods in Molecular Biology; Vol. 1999).

Research output: Chapter in Book/Report/Conference proceedingChapter

Pond, KW & Ellis, N 2019, Quantification of double-strand breaks in mammalian cells using pulsed-field gel electrophoresis. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1999, Humana Press Inc., pp. 75-85. https://doi.org/10.1007/978-1-4939-9500-4_4
Pond KW, Ellis N. Quantification of double-strand breaks in mammalian cells using pulsed-field gel electrophoresis. In Methods in Molecular Biology. Humana Press Inc. 2019. p. 75-85. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-9500-4_4
Pond, Kelvin W. ; Ellis, Nathan. / Quantification of double-strand breaks in mammalian cells using pulsed-field gel electrophoresis. Methods in Molecular Biology. Humana Press Inc., 2019. pp. 75-85 (Methods in Molecular Biology).
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