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. 2017:1562:45-53.
doi: 10.1007/978-1-4939-6807-7_4.

Genome-Wide Location Analyses of N6-Methyladenosine Modifications (m6A-Seq)

Benoit Molinie  1 , Cosmas C Giallourakis  2
Affiliations

Genome-Wide Location Analyses of N6-Methyladenosine Modifications (m6A-Seq)

Benoit Molinie et al. Methods Mol Biol. 2017.

Abstract

N6-methyladenosine-sequencing (m6A-seq) is a critical tool to obtain an unbiased genome-wide picture of m6A sites of modification at high resolution. It allows the study of the impact of various perturbations on m6A modification distribution and the study of m6A functions. Herein, we describe the m6A-seq protocol, which entails RNA immunoprecipitation (RIP) performed on fragmented poly(A) RNA utilizing anti-m6A antibodies. The captured/enriched m6A positive RNA fragments are subsequently sequenced by RNA-seq in parallel with background control non-immunoprecipitated input RNA fragments. Analyses reveal peaks of m6A enrichment containing sites of modifications analogous to chromatin modification immunoprecipitation experiments.

Keywords: Epitranscriptome; Genome-wide; METTL14; METTL3; N6-Methyladenosine; m6A-Seq.

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Figures

Fig 1
Fig 1
Work flow and schematic diagram of m6A-seq protocol
Fig 2
Fig 2
m6A-seq example data tracks. UCSC genome browser tracks of m6A-seq position analyses for two genes of our previously published H1-ESC data (red for m6A-RIP and gray for input)
See this image and copyright information in PMC

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