RiboMeth-seq: Profiling of 2'-O-Me in RNA

TY - CHAP

T1 - RiboMeth-seq

T2 - Profiling of 2'-O-Me in RNA

AU - Krogh, Nicolai

AU - Birkedal, Ulf

AU - Nielsen, Henrik

PY - 2017

Y1 - 2017

N2 - 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.

AB - 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.

U2 - 10.1007/978-1-4939-6807-7_13

DO - 10.1007/978-1-4939-6807-7_13

M3 - Book chapter

C2 - 28349462

SN - 978-1-4939-6805-3

VL - 1562

T3 - Methods in Molecular Biology

SP - 189

EP - 209

BT - RNA Methylation

PB - Springer

ER -