A single-nucleotide resolution sequencing method of N6-adenine methylation sites in DNA and RNA is described. Using sodium nitrite under acidic conditions, chemoselective deamination of unmethylated adenines readily occurs, without competing deamination of N6-adenine sites. The deamination of adenines results in the formation of hypoxanthine bases, which are read by polymerases and reverse transcriptases as guanine; the methylated adenine sites resist deamination and are read as adenine. The approach, when coupled with high-throughput DNA sequencing and mutational analysis, enables the identification of N6-adenine sites in RNA and DNA within various sequence contexts.
CITATION STYLE
Mahdavi-Amiri, Y., Chung Kim Chung, K., & Hili, R. (2021). Single-nucleotide resolution of: N 6-adenine methylation sites in DNA and RNA by nitrite sequencing. Chemical Science, 12(2), 606–612. https://doi.org/10.1039/d0sc03509b
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