Detection of mA in single cultured cells using scDART-seq.

Most techniques for mapping mA-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 mA 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 mA sites. For complete details on the use and execution of this protocol, please refer to Tegowski et al. (2022).