title: Detection and Quantification of N6-Methyladenosine in Messenger RNA by TLC.
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.
authors.affiliation: School of Biosciences, The University of Nottingham, Plant sciences, Sutton Bonington Campus, Sutton Bonington, Leicestershire, LE12 5RD, UK.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Bodi
authors.affiliation: School of Biosciences, The University of Nottingham, Plant sciences, Sutton Bonington Campus, Sutton Bonington, Leicestershire, LE12 5RD, UK. rupert.fray@nottingham.ac.uk.
authors.forename: Rupert G
authors.initials: RG
authors.lastname: Fray
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives*; D002855:Chromatography, Thin Layer / Q000379:methods; D044127:Epigenesis, Genetic; D006868:Hydrolysis; D007553:Isotope Labeling; D008745:Methylation; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*; D015719:Single-Strand Specific DNA and RNA Endonucleases; D059467:Transcriptome
chemical_list: D012333:RNA, Messenger; C010223:N-methyladenosine; D015719:Single-Strand Specific DNA and RNA Endonucleases; D000241:Adenosine
keywords: Epitranscriptome; N 6-Methyladenosine; TLC; Thin Layer Chromatography; m6A; mRNA Methylation