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High-Resolution Mapping of N6-Methyladenosine Using m6A Crosslinking Immunoprecipitation Sequencing (m6A-CLIP-Seq)

  • Phillip J. Hsu
  • Chuan HeEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1870)

Abstract

N6-Methyladenosine, an abundant chemical modification in mRNA, plays crucial roles in regulating gene expression and biological processes. Research on m6A and its functions has progressed rapidly in the past few years, aided substantially by advances in high-throughput sequencing-based methods to profile m6A along the transcriptome. We present here a protocol for m6A crosslinking immunoprecipitation sequencing (m6A-CLIP-seq), which profiles m6A on mRNA at high resolution from as little as 1 μg of poly(A)-selected mRNA.

Key words

N6-Methyladenosine Transcriptome Methylome Affinity purification Sequencing 

Notes

Acknowledgements

We thank Caroline Vissers, Boxuan Zhao, and Allen Zhu for helpful comments. P.J.H. is supported by the NIH Medical Scientist National Research Service Award T32 GM007281. C.H. is supported by National Institutes of Health grants GM071440 and HG008935. C.H. is an investigator of the Howard Hughes Medical Institute.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Chemistry and Institute for Biophysical Dynamics; Howard Hughes Medical InstituteThe University of ChicagoChicagoUSA
  2. 2.Medical Scientist Training Program/Committee on ImmunologyThe University of ChicagoChicagoUSA
  3. 3.Department of Biochemistry and Molecular BiologyThe University of ChicagoChicagoUSA

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