Stranded Whole Transcriptome RNA-Seq for All RNA Types

Curr Protoc Hum Genet. 2015 Jan 20:84:11.14.1-11.14.23. doi: 10.1002/0471142905.hg1114s84.

Abstract

Stranded whole transcriptome RNA-Seq described in this unit captures quantitative expression data for all types of RNA including, but not limited to, miRNA (microRNA), piRNA (Piwi-interacting RNA), snoRNA (small nucleolar RNA), lincRNA (large non-coding intergenic RNA), SRP RNA (signal recognition particle RNA), tRNA (transfer RNA), mtRNA (mitochondrial RNA), and mRNA (messenger RNA). The size and nature of these types of RNA are irrelevant to the approach described here. Barcoded libraries for multiplexing on the Illumina platform are generated with this approach but it can be applied to other platforms with a few modifications.

Keywords: RNA-Seq; duplex-specific nuclease; gene expression; transcriptome.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Complementary / analysis
  • DNA, Complementary / genetics
  • Gene Library
  • High-Throughput Nucleotide Sequencing / instrumentation
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Polymerase Chain Reaction / methods
  • RNA / analysis*
  • RNA / genetics
  • RNA, Long Noncoding / analysis*
  • RNA, Long Noncoding / genetics
  • RNA, Messenger / analysis*
  • RNA, Messenger / genetics
  • RNA, Mitochondrial
  • RNA, Small Untranslated / analysis*
  • RNA, Small Untranslated / classification
  • RNA, Small Untranslated / genetics
  • RNA, Transfer / analysis*
  • RNA, Transfer / genetics
  • Reverse Transcription
  • Transcriptome*

Substances

  • DNA, Complementary
  • RNA, Long Noncoding
  • RNA, Messenger
  • RNA, Mitochondrial
  • RNA, Small Untranslated
  • RNA
  • RNA, Transfer