Barcoded cDNA library preparation for small RNA profiling by next-generation sequencing

Methods. 2012 Oct;58(2):164-70. doi: 10.1016/j.ymeth.2012.07.030. Epub 2012 Aug 7.

Abstract

The characterization of post-transcriptional gene regulation by small regulatory (20-30 nt) RNAs, particularly miRNAs and piRNAs, has become a major focus of research in recent years. A prerequisite for characterizing small RNAs is their identification and quantification across different developmental stages, and in normal and disease tissues, as well as model cell lines. Here we present a step-by-step protocol for generating barcoded small RNA cDNA libraries compatible with Illumina HiSeq sequencing, thereby facilitating miRNA and other small RNA profiling of large sample collections.

Publication types

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

MeSH terms

  • Base Sequence
  • Gene Expression Profiling / methods
  • Gene Library*
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • MicroRNAs* / chemistry
  • MicroRNAs* / genetics
  • RNA, Small Interfering / chemistry
  • RNA, Small Interfering / genetics
  • Sequence Analysis, RNA / methods
  • Specimen Handling*

Substances

  • MicroRNAs
  • RNA, Small Interfering