Mapping yeast origins of replication via single-stranded DNA detection

Methods. 2007 Feb;41(2):151-7. doi: 10.1016/j.ymeth.2006.07.023.

Abstract

Studies in th Saccharomyces cerevisiae have provided a framework for understanding how eukaryotic cells replicate their chromosomal DNA to ensure faithful transmission of genetic information to their daughter cells. In particular, S. cerevisiae is the first eukaryote to have its origins of replication mapped on a genomic scale, by three independent groups using three different microarray-based approaches. Here we describe a new technique of origin mapping via detection of single-stranded DNA in yeast. This method not only identified the majority of previously discovered origins, but also detected new ones. We have also shown that this technique can identify origins in Schizosaccharomyces pombe, illustrating the utility of this method for origin mapping in other eukaryotes.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Chromosome Mapping / methods*
  • DNA, Single-Stranded / analysis*
  • DNA, Single-Stranded / genetics
  • Nucleic Acid Hybridization / methods
  • Oligonucleotide Array Sequence Analysis / methods*
  • Replication Origin*
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces / metabolism

Substances

  • DNA, Single-Stranded