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Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution
Published on: January 16, 2017
Methods for Extraction of RNA, Proteins, or Protein Complexes from Subcellular Compartments of Eukaryotic Cells
Kyu-Hyeon Yeom1, Andrey Damianov2
1Department of Microbiology, Immunology, and Molecular Genetics, The David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, 90095, USA. kyeom@microbio.ucla.edu.
Abstract:
Eukaryotic organelles or subcellular compartments can be selectively isolated based on their physical density and their stability in the presence of nonionic detergents. This chapter describes a protocol for the preparation of cytoplasm, nucleoplasm, and chromatin, in addition to isolation of RNA and proteins from these fractions. Proteins and protein complexes stably associated with chromatin and other high molecular weight nuclear components can be extracted under non-denaturing conditions by enzymatic digestion of RNA and DNA. The chapter also includes a detailed extraction protocol from highly purified nuclei.
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