Isolation of yeast nuclei and micrococcal nuclease mapping of nucleosome positioning.

Zhengjian Zhang, Joseph C. Reese

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

Chromatin structure and nucleosome positioning play a crucial role in gene expression regulation. Nucleosome positioning is often inferred by the protection of underlying DNA to nucleases. Because nucleases are excluded by plasma membranes, chromatin mapping requires isolating nuclei from cells and digesting the chromatin in situ with nucleases. The quality of this data is highly dependent on the nuclei preparation. Here we describe a method to isolate nuclei from the budding yeast Saccharomyces cerevisiae and the use of micrococcal nuclease to map the chromatin structure at the RNR3 gene. Nuclei isolated by this procedure are competent for many of the common chromatin mapping and detection procedures.

Original languageEnglish (US)
Pages (from-to)245-255
Number of pages11
JournalMethods in molecular biology (Clifton, N.J.)
Volume313
StatePublished - 2006

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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