6533b7d3fe1ef96bd1260a02

RESEARCH PRODUCT

Analysis of RNA modifications by liquid chromatography–tandem mass spectrometry

Kathrin ThüringKatharina SchmidPatrick KellerMark Helm

subject

0301 basic medicineChromatographyChemistrySelected reaction monitoringMs analysisRNATandem mass spectrometryMass spectrometryModified nucleosidesGeneral Biochemistry Genetics and Molecular BiologyHighly sensitive03 medical and health sciences030104 developmental biologyTandem Mass SpectrometryLiquid chromatography–mass spectrometryHumansRNARNA Processing Post-TranscriptionalMolecular BiologyChromatography Liquid

description

The analysis of RNA modifications is of high importance in order to address a wide range of biological questions. Therefore, a highly sensitive and accurate method such as liquid chromatography-tandem mass spectrometry (LC-MS/MS) has to be available. By using different LC-MS/MS procedures, it is not only possible to quantify very low amounts of RNA modifications, but also to detect probably unknown modified nucleosides. For these cases the dynamic multiple reaction monitoring and the neutral loss scan are the most common techniques. Here, we provide the whole workflow for analyzing RNA samples regarding their modification content. This includes an equipment list, the preparation of required solutions/enzymes and the creation of an internal standard or nucleoside stocks for internal or external calibration. Furthermore, we describe the preparation of RNA samples for the subsequent LC-MS/MS analysis and the corresponding analysis process.

yearjournalcountryeditionlanguage
2016-09-01Methods
https://doi.org/10.1016/j.ymeth.2016.03.019