Genome-Scale Mapping of DNase I Hypersensitivity

Genome-Scale Mapping of DNase I Hypersensitivity

Genome-Scale Mapping of DNase I Hypersensitivity

ABSTRACT

DNase I-seq is a global and high-resolution method that uses the nonspecific endonuclease
DNase I to map chromatin accessibility. These accessible regions, designated as
DNase I hypersensitive sites (DHSs), define the regulatory features, (e.g., promoters,
enhancers, insulators, and locus control regions) of complex genomes. In this unit, methods
are described for nuclei isolation, digestion of nuclei with limiting concentrations of
DNase I, and the biochemical fractionation of DNase I hypersensitive sites in preparation
for high-throughput sequencing. DNase I-seq is an unbiased and robust method that is
not predicated on an a priori understanding of regulatory patterns or chromatin features.
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