page_title: Epitranscriptomic sequencing
section_title: Methods for profiling N6-methyladenosine
hierarchical_section_title: Epitranscriptomic sequencing / Methods for profiling N6-methyladenosine
caption_attribution_description: English: Methods developed to profile m6A
context_page_description: In epitranscriptomic sequencing, most methods focus on either enrichment and purification of the modified RNA molecules before running on the RNA sequencer, or improving or modifying bioinformatics analysis pipelines to call the modification peaks. Most methods have been adapted and optimized for mRNA molecules, except for modified bisulfite sequencing for profiling 5-methylcytidine which was optimized for tRNAs and rRNAs.
  There are six major classes of chemical modifications found in RNA molecules: N⁶-methyladenosine, N6,2'-O-dimethyladenosine, 5-methylcytidine, 5-hydroxylmethylcytidine, inosine, and pseudouridine. Various sequencing methods have been developed to profile each type of modification. The scale, resolution, sensitivity, and limitations associated with each method and the corresponding bioinformatics tools used will be discussed.
context_section_description: Methylation of adenosine does not affect its ability to base-pair with thymidine or uracil, so  N⁶-methyladenosine (m⁶A) cannot be detected using standard sequencing or hybridization methods. This modification is marked by the methylation of the adenosine base at the nitrogen-6 position. It is abundantly found in polyA+ mRNA; also found in tRNA, rRNA, snRNA, and long ncRNA.
caption: Methods developed to profile N6-methyladenosine.
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