Double blocking gap-filling-ligation coupled with cascade isothermal amplification for ultrasensitive quantification of N6-methyladenosine

Abstract

We developed a method for quantifying N⁶-methyladenosine at one-nucleotide resolution based on double blocking gap-filling-ligation and cascade isothermal amplification. This proposed method can detect as low as 1 fM target RNA, achieving selectivity up to approximately 100-fold between m⁶A and A, and has been successfully applied to the analysis of m⁶A at specific sites in cell samples.