Accurate detection of m6A RNA modifications in native RNA sequences [Yeast]

The field of epitranscriptomics has undergone an enormous expansion in the last few years; however, a major limitation is the lack of generic methods to map RNA modifications transcriptome-wide. Here we show that using Oxford Nanopore Technologies, N6-methyladenosine (m6A) RNA modifications can be detected with high accuracy, in the form of systematic errors and decreased base-calling qualities. Our results open new avenues to investigate the universe of RNA modifications with single nucleotide resolution, in individual RNA molecules. Total RNA from meiotic SK1 yeast cells was extracted. Enrichment of polyadenylated RNA from total RNA was performed using Oligo(dT) dynabeads. Purified PolyA RNA was sequenced with Direct RNA Nanopore Sequencing. Three biological replicates were performed for each strain .