Dot Blot Analysis of N6-methyladenosine RNA Modification Levels

Lisha Shen; Zhe Liang; Hao Yu

doi:10.21769/BioProtoc.2095

Dot Blot Analysis of N⁶-methyladenosine RNA Modification Levels

Bio Protoc. 2017 Jan 5;7(1):e2095. doi: 10.21769/BioProtoc.2095.

Authors

Lisha Shen¹, Zhe Liang¹, Hao Yu¹

Affiliation

¹ Temasek Life Sciences Laboratory and Department of Biological Sciences, National University of Singapore, Singapore.

Abstract

N⁶-methyladenosine (m⁶A) is the most prevalent internal modification of eukaryotic messenger RNA (mRNA). The total amount of m⁶A can be detected by several methods, such as dot blot analysis using specific m⁶A antibodies and quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) ( Fu et al., 2014 ; Shen et al., 2016 ). Here we describe the method for fast detection of total m⁶A levels in mRNA by dot blot analysis using a specific m⁶A antibody.

Keywords: m6A; Dot blot; RNA modification.