Chapter title |
RiboMeth-seq: Profiling of 2′-O-Me in RNA
|
---|---|
Chapter number | 13 |
Book title |
RNA Methylation
|
Published in |
Methods in molecular biology, March 2017
|
DOI | 10.1007/978-1-4939-6807-7_13 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6805-3, 978-1-4939-6807-7
|
Authors |
Krogh, Nicolai, Birkedal, Ulf, Nielsen, Henrik, Nicolai Krogh, Ulf Birkedal, Henrik Nielsen |
Editors |
Alexandra Lusser |
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. |
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