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Analysis of SUC2 Promoter Structure by Nucleosome Scanning

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 809)

Abstract

Chromatin remodeling is a key mode of transcriptional regulation, and studying the nucleosome positioning at promoters is an important means to understand how genes are regulated. Nucleosome scanning is a convenient method to study nucleosome positioning. Yeast cells are converted to spheroplasts and nuclei are isolated. The nuclei are then digested by micrococcal nuclease to yield mononucleosome-sized DNA. Using a set of overlapping primers that cover the entire promoter, quantitative real-time PCR is performed using the mononucleosome DNA as the template. The nucleosome enrichment for each primer is calculated to yield a map of nucleosome occupancy across the promoter.

Key words

Saccharomyces cerevisiae Nucleosome scanning Chromatin Promoter structure Transcription PLC1 Inositol polyphosphates 
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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Cell BiologyNew York University School of MedicineNew YorkUSA
  2. 2.Department of Biological SciencesSt. John’s UniversityQueensUSA

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