Detecting repair intermediates in vivo: effects of DNA damage response genes on single-stranded DNA accumulation at uncapped telomeres in budding yeast

Steven Foster, Mikhajlo K. Zubko, Laura Maringele, David Lydall

    Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

    15 Citations (Scopus)

    Abstract

    Single-stranded DNA (ssDNA) is an important intermediate in many DNA repair pathways. Here we describe protocols that permit the measurement of ssDNA that has arisen in the yeast genome in vivo, in response to telomere uncapping. Yeast strains defective in DNA damage response (DDR) genes can be used to infer the roles of the corresponding proteins in regulating ssDNA production and in responding to ssDNA. Using column based methods to purify yeast genomic DNA and quantitative amplification of single-stranded DNA (QAOS) it is possible to measure ssDNA at numerous single copy loci in the yeast genome. We describe how to measure ssDNA in synchronous cultures of cdc13-1 mutants, containing a temperature sensitive mutation in an essential telomere capping protein, and in asynchronous cultures of yku70Delta mutants also defective in telomere capping.
    Original languageEnglish
    Title of host publicationEdit Methods in Enzymology
    Subtitle of host publicationDNA Repair, Part B, Volume 409
    EditorsJudith Campbell, Paul Modrich
    Chapter16
    Pages285-300
    Number of pages16
    Volume409
    ISBN (Electronic)9780080464671
    Publication statusPublished - 1 May 2006

    Publication series

    NameMethods in Enzymology
    ISSN (Print)0076-6879

    ASJC Scopus subject areas

    • General Biochemistry,Genetics and Molecular Biology

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