Quantitative analysis of m6A RNA modification by LC-MS

Lavina Mathur; Sunhee Jung; Cholsoon Jang; Gina Lee

STAR Protocols (Sep 2021)

Quantitative analysis of m6A RNA modification by LC-MS

Lavina Mathur,
Sunhee Jung,
Cholsoon Jang,
Gina Lee

Affiliations

Lavina Mathur: Department of Microbiology and Molecular Genetics, Chao Family Comprehensive Cancer Center, University of California Irvine School of Medicine, Irvine, CA 92697-4025, USA
Sunhee Jung: Department of Biological Chemistry, Chao Family Comprehensive Cancer Center, University of California Irvine School of Medicine, Irvine, CA 92697-4025, USA
Cholsoon Jang: Department of Biological Chemistry, Chao Family Comprehensive Cancer Center, University of California Irvine School of Medicine, Irvine, CA 92697-4025, USA; Corresponding author
Gina Lee: Department of Microbiology and Molecular Genetics, Chao Family Comprehensive Cancer Center, University of California Irvine School of Medicine, Irvine, CA 92697-4025, USA; Corresponding author

Journal volume & issue: Vol. 2, no. 3
p. 100724

Abstract

Read online

Summary: N6-adenosine methylation (m6A) of messenger RNA (mRNA) plays key regulatory roles in gene expression. Accurate measurement of m6A levels is thus critical to understand its dynamic changes in various biological settings. Here, we provide a protocol to quantitate the levels of adenosine and m6A in cellular mRNAs. Using nuclease and phosphatase, we digest mRNA into nucleosides, which are subsequently quantified using liquid chromatography mass spectrometry.For complete details on the use and execution of this protocol, please refer to Cho et al. (2021).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

About the journal

Abstract

Keywords