Molinie, Benoit, Giallourakis, Cosmas C., Benoit Molinie, Cosmas C. Giallourakis
Editors
Alexandra Lusser
Abstract
N(6)-methyladenosine-sequencing (m(6)A-seq) is a critical tool to obtain an unbiased genome-wide picture of m(6)A sites of modification at high resolution. It allows the study of the impact of various perturbations on m(6)A modification distribution and the study of m(6)A functions. Herein, we describe the m(6)A-seq protocol, which entails RNA immunoprecipitation (RIP) performed on fragmented poly(A) RNA utilizing anti-m(6)A antibodies. The captured/enriched m(6)A positive RNA fragments are subsequently sequenced by RNA-seq in parallel with background control non-immunoprecipitated input RNA fragments. Analyses reveal peaks of m(6)A enrichment containing sites of modifications analogous to chromatin modification immunoprecipitation experiments.
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