Double blocking gap-filling-ligation coupled with cascade isothermal amplification for ultrasensitive quantification of N-6-methyladenosine

We developed a method for quantifying N-6-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 m(6)A and A, and has been successfully applied to the analysis of m(6)A at specific sites in cell samples.

Automated summary

We have developed a method to quantify N-6-methyladenosine with high resolution using double blocking gap-filling-ligation and cascade isothermal amplification. This method can detect as low as 1 fM target RNA and achieve a selectivity of approximately 100-fold between m(6)A and A. It has been successfully applied to analyze m(6)A at specific sites in cell samples.