Relative Quantification of Residue-Specific m⁶A RNA Methylation Using m⁶A-RT-QPCR

Technological advances in high-throughput sequencing in combination with antibody enrichment and/or induced nucleotide-specific chemical modifications have accelerated the mapping of epitranscriptomic modifications. However, site-specific detection and quantification of m⁶A are still technically challenging. Here, we describe a simple RT-QPCR-based approach for the relative quantification of candidate m⁶A regions that takes advantage of the diminished capacity of BstI enzyme to retrotranscribe m⁶A residues.