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Probing RNA Structure with Dimethyl Sulfate Mutational Profiling with Sequencing In Vitro and in Cells
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Probing RNA Structure with Dimethyl Sulfate Mutational Profiling with Sequencing In Vitro and in Cells

Probing RNA Structure with Dimethyl Sulfate Mutational Profiling with Sequencing In Vitro and in Cells

DOI:
10.3791/64820-v

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10:34 min

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December 09, 2022

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,
1Department of Microbiology,Harvard Medical School

Chapters

  • 00:04Introduction
  • 00:36Gene‐Specific In Vitro DMS Modification
  • 02:49Gene‐Specific RT‐PCR of Modified RNA
  • 04:19Whole‐Genome DMS‐MaP Using Virus‐Infected Cells
  • 07:17Analysis of the Sequencing Data
  • 08:12Results: Probing RNA Structure with Dimethyl Sulfate Mutational Profiling and Sequencing
  • 10:00Conclusion

Summary

Automatic Translation

Automatic Translation

The protocol provides instruction for modifying RNA with dimethyl sulfate for mutational profiling experiments. It includes in vitro and in vivo probing with two alternative library preparation methods.

Tags

Keywords: RNA StructureDMS-MaPSequencingIn VitroIn CellsRNA Structure EnsembleBiological ResearchRefolding BufferDimethyl SulfateBeta-mercaptoethanolSpectrophotometerTGIRTComplementary DNAPCR

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