Quantification of m6A RNA modification in Caenorhabditis elegans by LC‐MS

Abstract N 6 ‐methyladenosine (m 6 A), one of the most abundant RNA modifications, has been found to be present in RNA in various life forms and plays a crucial regulatory role in gene expression. Caenorhabditis elegans ( C . elegans ) is a unique model organism with abundant m 6 A modifications. To analyze and understand the function of m 6 A in C . elegans , mass spectrometry is preferred because of its remarkable sensitivity and accuracy in detecting modified nucleosides. Here, we present a protocol for quantifying adenosine and m 6 A levels in C . elegans RNA. Using nucleases and phosphatases, we digested RNA into nucleosides and then quantified it using liquid chromatography‐mass spectrometry (LC–MS).