Deep sequencing of small chromatin-associated RNA: isolation and library preparation

Methods Mol Biol. 2014:1094:343-53. doi: 10.1007/978-1-62703-706-8_27.

Abstract

Chromatin-associated RNA (caRNA) is a newly identified class of RNA molecules stably associated with chromatin, maintaining the higher order structure of euchromatic regions in an accessible form (Schubert et al., Mol Cell, doi:10.1016/j.molcel.2012.08.021, 2012). This section provides a detailed protocol describing the isolation of small caRNA from Drosophila cells and preparation of libraries suited for stranded small (20-200 bp) RNA deep sequencing on the Illumina platform.

MeSH terms

  • Animals
  • Chromatin / metabolism*
  • Drosophila melanogaster / metabolism
  • Gene Library*
  • High-Throughput Nucleotide Sequencing / methods*
  • RNA, Small Nuclear / genetics*
  • RNA, Small Nuclear / isolation & purification*
  • Sequence Analysis, RNA / methods*

Substances

  • Chromatin
  • RNA, Small Nuclear