Quantitative analysis of RNA modifications.

Abstract

RNA modifications impact numerous cellular processes such as pre-mRNA splicing and protein synthesis. The elucidation of the mechanisms by which these modifications impact cellular processes necessitates the ability to both detect and quantify the presence of these modifications within RNA molecules. Here, we present a detailed procedure that allows the detection and quantification of RNA base modifications. This procedure involves a number of techniques, including oligonucleotide-affinity selection, site-specific cleavage and radiolabeling, nuclease digestion, and thin layer chromatography.