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Nucleic Acids Res. 2014 Feb;42(4):e27. doi: 10.1093/nar/gkt1160. Epub 2013 Nov 21.

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

Author information

1
Department of Chemistry, Lomonosov Moscow State University, Moscow 119992, Russia, Department of Bioinformatics and Bioengineering, Lomonosov Moscow State University, Moscow 119992, Russia and A.N. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119992, Russia.

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.

PMID:
24265225
PMCID:
PMC3936739
DOI:
10.1093/nar/gkt1160
[Indexed for MEDLINE]
Free PMC Article

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