m⁶A-label-seq: A metabolic labeling protocol to detect transcriptome-wide mRNA N⁶-methyladenosine (m⁶A) at base...

We describe here a metabolic labeling protocol for detecting cellular transcriptome-wide mRNA N -methyladenosine (m⁶A) at base resolution. By feeding cells an analog of methionine, potential mRNA m⁶A forming sites are replaced with the N -allyladenosine (a⁶A). A mild chemical iodination of a⁶A in RNA results in its opposite base misincorporation during RNA reverse transcription, and thus m⁶A locations could be precisely identified in the high-throughput sequencing. m⁶A-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.