Small RNA library cloning procedure for deep sequencing of specific endogenous siRNA classes in Caenorhabditis elegans

Methods Mol Biol. 2014:1173:59-70. doi: 10.1007/978-1-4939-0931-5_6.

Abstract

In recent years, distinct classes of small RNAs ranging in size from ~21 to 26 nucleotides have been discovered and shown to play important roles in a wide array of cellular functions. Because of the abundance of these small RNAs, library preparation from an RNA sample followed by deep sequencing provides the identity and quantity of a particular class of small RNAs. In this chapter we describe a detailed protocol for preparing small RNA libraries for deep sequencing on the Illumina platform from the nematode C. elegans.

MeSH terms

  • Animals
  • Base Sequence
  • Caenorhabditis elegans / genetics*
  • Cloning, Molecular / methods*
  • Gene Library
  • High-Throughput Nucleotide Sequencing / methods*
  • RNA, Small Interfering / chemistry
  • RNA, Small Interfering / genetics*
  • Reverse Transcription

Substances

  • RNA, Small Interfering