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Structural Analysis of RNA by Small-Angle X-ray Scattering

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RNA Spectroscopy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2113))

Abstract

Over the past two decades small-angle X-ray scattering (SAXS) has become a popular method to characterize solutions of biomolecules including ribonucleic acid (RNA). In an integrative structural approach, SAXS is complementary to crystallography, NMR, and electron microscopy and provides information about RNA architecture and dynamics. This chapter highlights the practical advantages of combining size-exclusion chromatography and SAXS at synchrotron facilities. It is illustrated by practical case studies of samples ranging from single hairpins and tRNA to a large IRES. The emphasis is also put on sample preparation which is a critical step of SAXS analysis and on optimized protocols for in vitro RNA synthesis ensuring the production of mg amount of pure and homogeneous molecules.

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Acknowledgments

The authors thank the team of the SWING beamline (SOLEIL synchrotron, Saint-Aubin, France), in particular Pierre Roblin and Javier Pérez, for beam time provision and assistance during data collection and processing, as well as José Teixeira (LLB, CEA Saclay, France) for his critical reading of the manuscript and fruitful discussion. The work described in this chapter was supported by the French Centre National de la Recherche Scientifique, the University of Strasbourg, the LabEx consortium “NetRNA” (ANR-10-LABX-0036_NETRNA).

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Authors and Affiliations

  1. Architecture et Réactivité de l’ARN, UPR 9002, IBMC, CNRS, Université de Strasbourg, Strasbourg, France

    Anne Théobald-Dietrich & Alexandra Bluhm

  2. Architecture et Réactivité de l’ARN - CNRS UPR 9002, Institut de Biologie Moléculaire et Cellulaire, Université de Strasbourg, Strasbourg, France

    Raphaël de Wijn, Kévin Rollet, Joëlle Rudinger-Thirion, Caroline Paulus, Bernard Lorber, Magali Frugier & Claude Sauter

  3. Synchrotron SOLEIL, Saint-Aubin, Gif-sur-Yvette, France

    Aurélien Thureau

Authors
  1. Anne Théobald-Dietrich
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  2. Raphaël de Wijn
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  3. Kévin Rollet
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  4. Alexandra Bluhm
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  5. Joëlle Rudinger-Thirion
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  6. Caroline Paulus
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  7. Bernard Lorber
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  8. Aurélien Thureau
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  9. Magali Frugier
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  10. Claude Sauter
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Corresponding author

Correspondence to Claude Sauter .

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Editors and Affiliations

  1. Université de Paris, Paris, France, Laboratoire Léon Brillouin LLB, CEA, CNRS UMR12, Université Paris Saclay, Gif-sur-Yvette, France

    Véronique Arluison

  2. Synchrotron SOLEIL L’Orme des Merisiers Saint Aubin, Gif-sur-Yvette, France

    Frank Wien

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Théobald-Dietrich, A. et al. (2020). Structural Analysis of RNA by Small-Angle X-ray Scattering. In: Arluison, V., Wien, F. (eds) RNA Spectroscopy. Methods in Molecular Biology, vol 2113. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0278-2_14

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  • DOI: https://doi.org/10.1007/978-1-0716-0278-2_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0277-5

  • Online ISBN: 978-1-0716-0278-2

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