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Issue 2, 2021
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Single-nucleotide resolution of N6-adenine methylation sites in DNA and RNA by nitrite sequencing

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Abstract

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.

Graphical abstract: Single-nucleotide resolution of N6-adenine methylation sites in DNA and RNA by nitrite sequencing

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Supplementary files

Article information


Submitted
24 Jun 2020
Accepted
04 Nov 2020
First published
05 Nov 2020

This article is Open Access
All publication charges for this article have been paid for by the Royal Society of Chemistry

Chem. Sci., 2021,12, 606-612
Article type
Edge Article

Single-nucleotide resolution of N6-adenine methylation sites in DNA and RNA by nitrite sequencing

Y. Mahdavi-Amiri, K. Chung Kim Chung and R. Hili, Chem. Sci., 2021, 12, 606
DOI: 10.1039/D0SC03509B

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