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Detection and Quantification of N6-Methyladenosine in Messenger RNA by TLC

  • Zsuzsanna Bodi
  • Rupert G. FrayEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1562)

Abstract

The base-modified nucleotide, N6-methyladenosine, is a relatively abundant modification found in the mRNA of most higher eukaryotes. Methylation levels can change dependent upon environmental conditions, cell differentiation state, or following knockdown of members of the methylase complex, and it is often useful to directly measure and compare N6-methyladenosine levels between samples. Two dimensional chromatography of radiolabeled nucleotides, following specific nuclease treatments, provides a robust, sensitive, and reproducible assay for this modification.

Key words

N6-Methyladenosine m6Thin Layer Chromatography TLC mRNA Methylation Epitranscriptome 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.School of BiosciencesThe University of NottinghamLeicestershireUK

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