Detection of m⁶A in single cultured cells using scDART-seq

Most techniques for mapping m⁶A-methylated RNAs transcriptome-wide require large amounts of RNA and have been limited to bulk cells and tissues. Here, we provide a detailed protocol for the identification of m⁶A sites in single HEK293T cells using single-cell DART-seq (scDART-seq). The protocol details how to generate cell lines with inducible expression of the APOBEC1-YTH transgene and the use of important controls for minimizing false positives. We also describe the bioinformatic analysis to identify m⁶A sites. For complete details on the use and execution of this protocol, please refer to Tegowski et al. (2022).