Literature DB >> 28349453

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

Benoit Molinie1, Cosmas C Giallourakis2.   

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.

Entities:  

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

Mesh:

Substances:

Year:  2017        PMID: 28349453      PMCID: PMC5755608          DOI: 10.1007/978-1-4939-6807-7_4

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN: 1064-3745


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