Detection of 5-formylcytosine in Mitochondrial Transcriptome

Posttranscriptional RNA modifications have recently emerged as essential posttranscriptional regulators of gene expression. Here we present two methods for single nucleotide resolution detection of 5-formylcytosine (fC) in RNA. The first relies on chemical protection of fC against bisulfite treatment, the second method is based on chemical reduction of fC to hmC. In combination with regular bisulfite treatment of RNA, the methods allow for precise mapping of fC. The protocol is used for fC detection in mtDNA-encoded RNA, however, it can be straightforwardly applied for transcriptome-wide analyses.