m(6)A-label-seq: A metabolic labeling protocol to detect transcriptome-wide mRNA N(6)-methyladenosine (m(6)A) at base resolution

We describe here a metabolic labeling protocol for detecting cellular transcriptome-wide mRNA N(6)-methyladenosine (m(6)A) at base resolution. By feeding cells an analog of methionine, potential mRNA m(6)A forming sites are replaced with the N(6)-allyladenosine (a(6)A). A mild chemical iodination of a(6)A in RNA results in its opposite base misincorporation during RNA reverse transcription, and thus m(6)A locations could be precisely identified in the high-throughput sequencing. m(6)A-label-seq provides a strategy to label and identify cellular epitranscriptomic modification sites. For complete details on the use and execution of this profile, please refer to Shu et al., 2020.