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LC-MS Analysis of Methylated RNA

  • Kathrin Thüring
  • Katharina Schmid
  • Patrick Keller
  • Mark HelmEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1562)

Abstract

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.

Key words

RNA methylation LC-MS DMRM NLS SIL-IS External calibration 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Kathrin Thüring
    • 1
  • Katharina Schmid
    • 1
  • Patrick Keller
    • 1
  • Mark Helm
    • 1
    Email author
  1. 1.Institute of Pharmacy and BiochemistryJohannes Gutenberg-University of MainzMainzGermany

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