Transcriptome-Wide Mapping of N6-Methyladenosine by m6A-Seq

Dan Dominissini, Sharon Moshitch-Moshkovitz, Ninette Amariglio, Gideon Rechavi

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

A detailed protocol for isolation and sequencing of an enriched population of m6A-methylated RNA fragments to create m6A methylome maps is outlined. Our approach was developed to fill a void that existed because of a lack of methods for the detection of m6A in RNA in an unbiased, high-throughput, and high-resolution manner. This method integrates immunoprecipitation of methylated, randomly fragmented RNA using a highly specific anti-m6A antibody to obtain an enriched population of modified fragments and massively parallel sequencing, resulting in mapping of this modification throughout the transcriptome.

Original languageEnglish
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages131-147
Number of pages17
DOIs
StatePublished - 11 Aug 2015

Publication series

NameMethods in Enzymology
Volume560
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Immunocapturing
  • N-Methyladenosine
  • Next-generation sequencing
  • RNA immunoprecipitation
  • RNA methylation

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