Ribosomal RNA depletion for efficient use of RNA-seq capacity

Curr Protoc Mol Biol. 2013 Jul:Chapter 4:Unit 4.19. doi: 10.1002/0471142727.mb0419s103.

Abstract

Ribosomal RNA (rRNA) is the most highly abundant component of RNA, comprising the majority (>80% to 90%) of the molecules present in a total RNA sample. Depletion of this rRNA fraction is desirable prior to performing an RNA-seq reaction, so that sequencing capacity can be focused on more informative parts of the transcriptome. This unit describes an rRNA depletion method based on selective hybridization of oligonucleotides to rRNA, recognition with a hybrid-specific antibody, and removal of the antibody-hybrid complex on magnetic beads.

MeSH terms

  • Antibodies / metabolism
  • High-Throughput Nucleotide Sequencing / methods*
  • Nucleic Acid Hybridization / methods
  • Oligonucleotides / genetics
  • Oligonucleotides / metabolism
  • RNA / genetics*
  • RNA / isolation & purification*
  • RNA, Ribosomal / genetics
  • RNA, Ribosomal / isolation & purification*
  • RNA, Ribosomal / metabolism
  • Specimen Handling / methods*

Substances

  • Antibodies
  • Oligonucleotides
  • RNA, Ribosomal
  • RNA