Nucleotide-Level Profiling of m⁵C RNA Methylation.

Mapping the position and quantifying the level of 5-methylcytosine (m(5)C) as a modification in different types of cellular RNA is an important objective in the emerging field of epitranscriptomics. Bisulfite conversion has long been the gold standard for detection of m(5)C in DNA but it can also be applied to RNA. Here, we detail methods for bisulfite treatment of RNA, locus-specific PCR amplification and detection of candidate sites by sequencing on the Illumina MiSeq platform.