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Practical Aspects of Sample Preparation and Setup of 1H R1ρ Relaxation Dispersion Experiments of RNA
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Practical Aspects of Sample Preparation and Setup of 1H R1ρ Relaxation Dispersion Experiments of RNA

Practical Aspects of Sample Preparation and Setup of 1H R1ρ Relaxation Dispersion Experiments of RNA

DOI:
10.3791/62470-v

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08:17 min

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July 09, 2021

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, ,
1Department of Medical Biochemistry and Biophysics,Karolinska Institutet

Chapters

  • 00:04Introduction
  • 00:34NMR Sample Preparation
  • 03:481H R1ρ Relaxation Dispersion
  • 05:29Results: Analysis of Produced Sample and Two Different Constructs Based on an RNA Hairpin
  • 07:39Conclusion

Summary

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We present a protocol to measure micro- to millisecond dynamics on 13C/15N-labeled and unlabeled RNA with 1H R1ρ relaxation dispersion nuclear magnetic resonance (NMR) spectroscopy. The focus of this protocol lies in high-purity sample preparation and setup of NMR experiments.

Tags

Keywords: RNA Sample PreparationNMR SetupProton R1ρ Relaxation DispersionConformational ExchangeIn Vitro TranscriptionRNase H CleavageHPLC PurificationNMR Tube CleaningD2O Addition

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