m6A-SAC-seq for quantitative whole transcriptome m6A profiling

This protocol describes a method for base-resolution, quantitative m6A sequencing in the whole transcriptome. The method generates highly reproducible results containing 31,233-129,263 sites using as low as 2 ng of poly-A RNA. The RNA is treated with recombinant MjDim1 enzyme with allylic-SAM cofactor under optimized conditions to selectively convert m6A residues to a6m6A, followed by adding I2 to convert am6A to EA. Reverse transcription induces mutations at EA sites, enabling identification of the sites labeled with a6m6A.