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Related Concept Videos

RNA-seq03:21

RNA-seq

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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
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Updated: Oct 17, 2025

A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues
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m6Am RNA modification detection by m6Am-seq.

Meiling Zhang1, Hanxiao Sun1, Kai Li2

  • 1State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China.

Methods (San Diego, Calif.)
|October 8, 2021
PubMed
Summary

We developed m6Am-seq, a new method to detect N6, 2'-O-dimethyladenosine (m6Am) modifications in RNA. This technique offers single-base resolution for studying m6Am and 5'-UTR m6A methylomes.

Keywords:
DemethylationRNA immunoprecipitationSingle-base resolutionm(6)Am

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Area of Science:

  • Molecular Biology
  • Epigenetics
  • RNA Biology

Background:

  • Dynamic RNA modifications regulate gene expression.
  • N6, 2 -O-dimethyladenosine (m6Am) is a key mRNA cap modification with diverse biological roles.
  • Current m6Am detection methods have limitations, hindering applications.

Purpose of the Study:

  • To develop a specific and sensitive method for detecting m6Am at single-base resolution.
  • To enable comprehensive analysis of the m6Am and 5 -UTR m6A methylomes.
  • To facilitate further functional and mechanistic studies of m6Am modification.

Main Methods:

  • m6Am-seq, a novel method combining optimized in-vitro demethylation assay and RNA immunoprecipitation.
  • Single-base resolution detection of m6Am.
  • Distinguishing m6Am from 5 -UTR m6A.

Main Results:

  • m6Am-seq accurately detects m6Am at single-base resolution.
  • The method can differentiate between m6Am and 5 -UTR m6A.
  • A detailed protocol for m6Am-seq, including experimental procedures and data analysis, is provided.

Conclusions:

  • m6Am-seq is a robust tool for mapping the m6Am and 5 -UTR m6A methylomes.
  • This method overcomes limitations of previous m6Am detection techniques.
  • Enables deeper understanding of m6Am's role in gene regulation.