Transcriptome-Wide Mapping of N6-Methyladenosine by m6A-Seq

A detailed protocol for isolation and sequencing of an enriched population of m(6)A-methylated RNA fragments to create m(6)A methylome maps is outlined. Our approach was developed to fill a void that existed because of a lack of methods for the detection of m(6)A in RNA in an unbiased, high-throughput, and high-resolution manner. This method integrates immunoprecipitation of methylated, randomly fragmented RNA using a highly specific anti-m(6)A antibody to obtain an enriched population of modified fragments and massively parallel sequencing, resulting in mapping of this modification throughout the transcriptome.