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Large Scale Purification of RNA Nanoparticles by Preparative Ultracentrifugation

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RNA Nanotechnology and Therapeutics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1297))

Abstract

Purification of large quantities of supramolecular RNA complexes is of paramount importance due to the large quantities of RNA needed and the purity requirements for in vitro and in vivo assays. Purification is generally carried out by liquid chromatography (HPLC), polyacrylamide gel electrophoresis (PAGE), or agarose gel electrophoresis (AGE). Here, we describe an efficient method for the large-scale purification of RNA prepared by in vitro transcription using T7 RNA polymerase by cesium chloride (CsCl) equilibrium density gradient ultracentrifugation and the large-scale purification of RNA nanoparticles by sucrose gradient rate-zonal ultracentrifugation or cushioned sucrose gradient rate-zonal ultracentrifugation.

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Acknowledgements

The research was supported by NIH grants R01-EB003730 and U01-CA151648 to P.G. The content is solely the responsibility of the authors and does not necessarily represent the official views of NIH. Funding to Peixuan Guo’s Endowed Chair in Nanobiotechnology position is from the William Fairish Endowment Fund. PG is a cofounder of Kylin Therapeutics, Inc., RNA Nano, LLC., and Biomotor and Nucleic Acid Nanotechnology Development Corp., Ltd.

Author information

Authors and Affiliations

  1. Nanobiotechnology Center, Markey Cancer Center, Department of Pharmaceutical Sciences, University of Kentucky, 789 S. Limestone Avenue, Room #565, Lexington, KY, 40536-0596, USA

    Daniel L. Jasinski, Farzin Haque & Peixuan Guo Ph.D.

  2. Beckman Coulter, Inc., 5350 Lakeview Dr., Indianapolis, IN, 46268, USA

    Chad T. Schwartz

Authors
  1. Daniel L. Jasinski
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  2. Chad T. Schwartz
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  3. Farzin Haque
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  4. Peixuan Guo Ph.D.
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Corresponding author

Correspondence to Peixuan Guo Ph.D. .

Editor information

Editors and Affiliations

  1. Nanobiotechnology Center, Markey Cancer Center, Department of Pharmaceutical Sciences, University of Kentucky, Lexington, Kentucky, USA

    Peixuan Guo

  2. Nanobiotechnology Center, Markey Cancer Center, Department of Pharmaceutical Sciences, University of Kentucky, Lexington, Kentucky, USA

    Farzin Haque

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Jasinski, D.L., Schwartz, C.T., Haque, F., Guo, P. (2015). Large Scale Purification of RNA Nanoparticles by Preparative Ultracentrifugation. In: Guo, P., Haque, F. (eds) RNA Nanotechnology and Therapeutics. Methods in Molecular Biology, vol 1297. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2562-9_5

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  • DOI: https://doi.org/10.1007/978-1-4939-2562-9_5

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2561-2

  • Online ISBN: 978-1-4939-2562-9

  • eBook Packages: Springer Protocols

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