The detection and measurement of base and nucleotide excision repair in cell-free extracts of the yeast saccharomyces cerevisiae

Zhigang Wang, Xiaohua Wu, Errol C. Friedberg

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

Excision repair of DNA is a major cellular mechanism to correct DNA lesions produced by a variety of physical and chemical agents. The availability of a cell-free system is expected to provide a powerful tool with which to study the complex biochemical pathways of excision repair in eukaryotes. Here we describe such a cell-free system in the yeast Saccharomyces cerevisiae that supports both nucleotide excision repair (NER) and base excision repair (BER) in vitro. Detailed methods and protocols are provided for the preparation of yeast cell-free extracts, damaged DNA substrates, in vitro NER assays, and in vitro BER assays, with an emphasis on technical aspects. NER and BER detected in this cell-free system apparently reflect the complete repair pathways.

Original languageEnglish (US)
Pages (from-to)177-186
Number of pages10
JournalMethods
Volume7
Issue number2
DOIs
StatePublished - Apr 1995

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

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