title: RiboMeth-seq: Profiling of 2'-O-Me in RNA.
abstract: RiboMeth-seq is a sequencing-based method for mapping and quantitation of one of the most abundant RNA modifications, ribose methylation. It is based on a simple chemical principle, namely the several orders of magnitude difference in nucleophilicity of a 2'-OH and a 2'-O-Me. Thus, the method combines alkaline fragmentation and a specialized library construction protocol based on 5'-OH and 2',3' cyclic phosphate ends to prepare RNA for sequencing. The read-ends of library fragments are used for mapping with nucleotide resolution and calculation of the fraction of molecules methylated at the 2'-O-Me sites.
authors.affiliation: Department of Cellular and Molecular Medicine, The Panum Institute, University of Copenhagen, 3 Blegdamsvej, 18.2.22, DK-2200N, Copenhagen, Denmark.
authors.forename: Nicolai
authors.initials: N
authors.lastname: Krogh
authors.affiliation: Department of Cellular and Molecular Medicine, The Panum Institute, University of Copenhagen, 3 Blegdamsvej, 18.2.22, DK-2200N, Copenhagen, Denmark.
authors.forename: Ulf
authors.initials: U
authors.lastname: Birkedal
authors.affiliation: Department of Cellular and Molecular Medicine, The Panum Institute, University of Copenhagen, 3 Blegdamsvej, 18.2.22, DK-2200N, Copenhagen, Denmark. hamra@sund.ku.dk.
authors.forename: Henrik
authors.initials: H
authors.lastname: Nielsen
mesh_terms: D019295:Computational Biology / Q000379:methods; D020869:Gene Expression Profiling; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing; D006868:Hydrolysis; D008745:Methylation; D020871:RNA Stability; D012335:RNA, Ribosomal / Q000737:chemistry / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D012266:Ribose / Q000737:chemistry; D017423:Sequence Analysis, RNA / Q000379:methods; D013223:Statistics as Topic / Q000379:methods; D059467:Transcriptome
chemical_list: D012335:RNA, Ribosomal; D012266:Ribose
keywords: 2′-O-Me; Box C/D snoRNA; RNA library; RNA-seq; Ribose methylation; Ribosomal RNA; Ribosome