RiboMeth-seq: Profiling of 2′-O-Me in RNA
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.
Key words2′-O-Me Ribose methylation Box C/D snoRNA Ribosome Ribosomal RNA RNA library RNA-seq
The work was supported by the Danish Research Council for Independent Research and the Danish Cancer Research Foundation.
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