Biochemical Analysis of D-Loop Extension and DNA Strand Displacement Synthesis

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

Abstract

Homologous recombination is a conserved genome maintenance pathway through which DNA double-strand breaks are eliminated and perturbed DNA replication forks and eroded telomeres are restored. The central step in homologous recombination is homology-dependent pairing between a single-stranded DNA tail with an intact duplex molecule to generate a displacement-loop (D-loop), followed by DNA synthesis within the D-loop platform. This chapter describes biochemical assays for (1) D-loop formation and DNA synthesis within the D-loop and (2) DNA strand displacement synthesis to test the role of DNA helicases (e.g., Pif1) in repair DNA synthesis. These mechanistic assays are valuable for elucidating the molecular details of HR.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages87-99
Number of pages13
DOIs
StatePublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2153
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • D-loop
  • DNA helicase
  • DNA polymerase
  • DNA synthesis
  • Homologous recombination
  • Recombinase

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology

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