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

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7.2K Views

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08:12 min

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January 3rd, 2019

DOI :

10.3791/58697-v

January 3rd, 2019

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Aleksandra Skrajna¹, Xiao-Cui Yang¹, Zbigniew Dominski¹^,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

Podsumowanie

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Single step Purification

Macromolecular Complexes

RNA Protein Complexes

Mass Spectrometry

RNA Binding Sites

Biotin

Photo cleavable Linker

Nuclear Extract

Streptavidin Agarose Beads

Adaptor PCB Oligonucleotide

Supernatant Collection

Binding Buffer

Centrifugation Process

Immobilization

Lab Protocol

Rozdziały w tym wideo

0:04

Title

0:51

Substrate Preparation

1:28

Complex Assembly

2:14

Immobilization of the RNA/protein Complexes on Streptavidin Beads

4:00

UV-Elution

5:08

Sample Analysis by Silver Staining Followed by Mass Spectrometry

5:59

Results: Analysis of UV-eluted Complexes

7:36

Conclusion

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