Analysis of SUC2 promoter structure by nucleosome scanning.

Jennifer Chang, Ales Vancura

Department of Cell Biology, New York University School of Medicine, New York, NY, USA.

Methods in molecular biology (Clifton, N.J.) 2012

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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.

Citation

Jennifer Chang, Ales Vancura. Analysis of SUC2 promoter structure by nucleosome scanning. Methods in molecular biology (Clifton, N.J.). 2012;809:321-33