Cleavage under targets and release using nuclease (CUT & RUN) is an innovative method to profile histone modifications and chromatin-bound proteins genome-wide. CUT & RUN offers two distinct advantages of requiring much fewer cells and providing strong signal-to-noise ratios in deep-sequencing data. Here, we describe a workflow starting from dissociation and sorting of mouse embryonic brains, CUT & RUN, and DNA library preparation to deep sequencing. With our workflow, researchers can obtain high-quality sequencing data to profile histones and chromatin-associated proteins by using as few as 100,000 neural progenitor cells (NPCs).
Keywords: CUT & RUN; DNA library prep; Drosophila spike-in; Epigenetics; Fluorescence-activated cell sorting (FACS); Genome-wide chromatin profiling; Histone modifications; Next-generation sequencing (NGS); Papain dissociation; Primary neural progenitor cells (NPCs); pAG-MNase.
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