UNIT 21.25 Capture Hybridization Analysis of RNA Targets (CHART). Issue 1 (January 2013)

• Record Type: Journal Article
• Authors: Simon, Matthew D.
• Is Part Of: Current protocols in molecular biology. Volume 101:Issue 1(2013)
• Page Start: 21.25.1
• Page End: 21.25.16
• Publication Date: 2013-01
• Subjects: CHART -- chromatin -- non‐coding RNA -- lncRNA; Molecular biology -- Technique -- Periodicals; Molecular biology -- Laboratory manuals; Molecular Biology -- methods; Biologie moléculaire -- Technique; Biologie moléculaire -- Manuels de laboratoire; Molecular biology; Molecular biology -- Technique; Laboratory Manual; Electronic reference sources; Laboratory manuals; 572.8028
• DOI: 10.1002/0471142727.mb2125s101
• Languages: English
• ISSNs: 1934-3639

Abstract

Abstract: The genome is regulated by trans ‐acting factors that bind to specific loci in chromatin. In addition to protein factors, it has become clear that large non‐coding RNAs can also act on chromatin at sites distant from where they are transcribed. This unit describes a means of identifying the genomic targets of those large non‐coding RNAs. To accomplish this, the endogenous RNA of interest (here Drosophila roX2 is used as an example) is enriched from cross‐linked chromatin extracts using short biotinylated complementary oligodeoxyribonucleotides. The targets of the RNA can be determined by examining the proteins and DNA that are enriched under these conditions. This analysis can be extended genome‐wide by subjecting the enriched DNA to deep sequencing. Curr. Protoc. Mol. Biol. 101:21.25.1–21.25.16. © 2013 by John Wiley & Sons, Inc.