6533b7d8fe1ef96bd126ab8b

RESEARCH PRODUCT

LC-MS Analysis of Methylated RNA

Kathrin ThüringKatharina SchmidMark HelmPatrick Keller

subject

0301 basic medicineChemistryRNA methylationSelected reaction monitoringRNA03 medical and health sciences030104 developmental biology0302 clinical medicineBiochemistryLiquid chromatography–mass spectrometryEnzymatic hydrolysisGene expressionMethylated DNA immunoprecipitationEpigenetics030217 neurology & neurosurgery

description

The detection and quantification of methylated RNA can be beneficial to understand certain cellular regulation processes such as transcriptional modulation of gene expression, immune response, or epigenetic alterations. Therefore, it is necessary to have methods available, which are extremely sensitive and accurate, for instance liquid chromatography-tandem mass spectrometry (LC-MS/MS). Here, we describe the preparation of RNA samples by enzymatic hydrolysis and the subsequent analysis of ribonucleosides by LC-MS/MS via NLS (Neutral loss scan) and DMRM (Dynamic multiple reaction monitoring). Also, we provide variations of these methods including chromatographic techniques and different kinds of quantification.

https://doi.org/10.1007/978-1-4939-6807-7_1