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Single-step Purification of Macromolecular Complexes Using RNA Attached to Biotin and a Photo-cleavable Linker
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Biochemistry
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JoVE Journal Biochemistry
Single-step Purification of Macromolecular Complexes Using RNA Attached to Biotin and a Photo-cleavable Linker

Single-step Purification of Macromolecular Complexes Using RNA Attached to Biotin and a Photo-cleavable Linker

English

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7,068 Views

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08:12 min

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January 03, 2019

DOI:

10.3791/58697-v

DOI:
10.3791/58697-v

•

08:12 min
January 03, 2019

•

3 Views
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1Integrative Program for Biological and Genome Sciences,University of North Carolina at Chapel Hill, 2Department of Biochemistry and Biophysics,University of North Carolina at Chapel Hill

Summary

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RNA/protein complexes purified using botin-streptavidin strategy are eluted to solution under denaturing conditions in a form unsuitable for further purification and functional analysis. Here, we describe a modification of this strategy that utilizes a photo-cleavable linker in RNA and a gentle UV-elution step, yielding native and fully functional RNA/protein complexes.

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Cite this Article

Skrajna, A., Yang, X., Dominski, Z. Single-step Purification of Macromolecular Complexes Using RNA Attached to Biotin and a Photo-cleavable Linker. J. Vis. Exp. (143), e58697, doi:10.3791/58697 (2019).

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