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Biochemistry

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

Article DOI: 10.3791/58697-v • 08:12 min • January 3rd, 2019
January 3rd, 2019 •
1, 1, 1,2
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

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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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Biochimica numero 143 purificazione di affinità complessi di RNA/proteine biotina streptavidina foto-spaccabili linker UV-eluizione U7 snRNP 3' fine lavorazione
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