title: Transcriptome-Wide Mapping of N⁶-Methyladenosine by m⁶A-Seq.
abstract: A detailed protocol for isolation and sequencing of an enriched population of m(6)A-methylated RNA fragments to create m(6)A methylome maps is outlined. Our approach was developed to fill a void that existed because of a lack of methods for the detection of m(6)A in RNA in an unbiased, high-throughput, and high-resolution manner. This method integrates immunoprecipitation of methylated, randomly fragmented RNA using a highly specific anti-m(6)A antibody to obtain an enriched population of modified fragments and massively parallel sequencing, resulting in mapping of this modification throughout the transcriptome.
authors.affiliation: Department of Chemistry, The University of Chicago, Chicago, Illinois, USA; Institute for Biophysical Dynamics, The University of Chicago, Chicago, Illinois, USA.
authors.forename: Dan
authors.initials: D
authors.lastname: Dominissini
authors.affiliation: Cancer Research Center, Chaim Sheba Medical Center, Tel Hashomer, Israel; Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Sharon
authors.initials: S
authors.lastname: Moshitch-Moshkovitz
authors.affiliation: Cancer Research Center, Chaim Sheba Medical Center, Tel Hashomer, Israel; Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Ninette
authors.initials: N
authors.lastname: Amariglio
authors.affiliation: Cancer Research Center, Chaim Sheba Medical Center, Tel Hashomer, Israel; Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel. Electronic address: gidi.rechavi@sheba.health.gov.il.
authors.forename: Gideon
authors.initials: G
authors.lastname: Rechavi
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D008745:Methylation; D012313:RNA / Q000737:chemistry* / Q000235:genetics; D059467:Transcriptome / Q000235:genetics*
chemical_list: D012313:RNA; C010223:N-methyladenosine; D000241:Adenosine
keywords: Immunocapturing; N(6)-Methyladenosine; Next-generation sequencing; RNA immunoprecipitation; RNA methylation