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Single-step purificazione di complessi macromolecolari usando RNA attaccato alla biotina e un Linker foto-spaccabili
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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
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Single-step purificazione di complessi macromolecolari usando RNA attaccato alla biotina e un Linker foto-spaccabili

italiano

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7,057 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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Complessi RNA/proteici purificati mediante botin-streptavidina strategia sono eluiti per soluzione in condizioni denaturanti in forma inadatta per ulteriore purificazione e analisi funzionale. Qui, descriviamo una modifica di questa strategia che utilizza un linker foto-spaccabili in RNA e un delicato passaggio di UV-eluizione, producendo complessi RNA/proteine native e completamente funzionale.

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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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