Method for site-specific detection of m6A nucleoside presence in RNA based on high-resolution melting (HRM) analysis

基于高分辨率熔解 (HRM) 分析的 RNA 中 m6A 核苷位点特异性检测方法

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作者:Anna Y Golovina, Margarita M Dzama, Kirill S Petriukov, Timofei S Zatsepin, Petr V Sergiev, Alexey A Bogdanov, Olga A Dontsova

Abstract

Chemical landscape of natural RNA species is decorated with the large number of modified nucleosides. Some of those could easily be detected by reverse transcription, while others permit only high-performance liquid chromatography or mass-spectrometry detection. Presence of m(6)A nucleoside at a particular position of long RNA molecule is challenging to observe. Here we report an easy and high-throughput method for detection of m(6)A nucleosides in RNA based on high-resolution melting analysis. The method relies on the previous knowledge of the modified nucleoside position at a particular place of RNA and allows rapid screening for conditions or genes necessary for formation of that modification.

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