RiboMeth-seq: Profiling of 2'-O-Me in RNA

Methods Mol Biol. 2017:1562:189-209. doi: 10.1007/978-1-4939-6807-7_13.

Abstract

RiboMeth-seq is a sequencing-based method for mapping and quantitation of one of the most abundant RNA modifications, ribose methylation. It is based on a simple chemical principle, namely the several orders of magnitude difference in nucleophilicity of a 2'-OH and a 2'-O-Me. Thus, the method combines alkaline fragmentation and a specialized library construction protocol based on 5'-OH and 2',3' cyclic phosphate ends to prepare RNA for sequencing. The read-ends of library fragments are used for mapping with nucleotide resolution and calculation of the fraction of molecules methylated at the 2'-O-Me sites.

Keywords: 2′-O-Me; Box C/D snoRNA; RNA library; RNA-seq; Ribose methylation; Ribosomal RNA; Ribosome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Computational Biology / methods
  • Gene Expression Profiling*
  • Gene Library
  • High-Throughput Nucleotide Sequencing*
  • Hydrolysis
  • Methylation
  • RNA Stability
  • RNA, Ribosomal / chemistry
  • RNA, Ribosomal / genetics*
  • Real-Time Polymerase Chain Reaction
  • Ribose / chemistry
  • Sequence Analysis, RNA* / methods
  • Statistics as Topic / methods
  • Transcriptome*

Substances

  • RNA, Ribosomal
  • Ribose