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Elektroforetisk mobilitet (EMSA) for Studiet af RNA-protein interaktioner: Den IRE / IRP Eksempel
JoVE Journal
Biology
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JoVE Journal Biology
Electrophoretic Mobility Shift Assay (EMSA) for the Study of RNA-Protein Interactions: The IRE/IRP Example
DOI:

12:44 min

December 03, 2014

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Chapters

  • 00:05Title
  • 01:28Preparation of Protein Extracts from Cultured Cells
  • 03:05Preparation of Protein Extracts from Mouse Liver and Spleen
  • 04:56Synthesis of Radiolabeled IRE-containing RNA Probe
  • 05:55Purification of Radiolabeled IRE-containing RNA Probe
  • 07:48Electrophoretic Mobility Shift Assay (EMSA)
  • 10:27Results: The Iron-dependence of IRE/IRP Interactions
  • 12:17Conclusion

Summary

Automatic Translation

Her præsenterer vi en protokol til at analysere RNA / protein-interaktioner. Den elektroforetiske mobilitet (EMSA) er baseret på forskellen migration af RNA / protein-komplekser og fri RNA under nativ gelelektroforese. Ved anvendelse af en radioaktivt mærket RNA-probe, kan RNA / protein komplekser visualiseret ved autoradiografi.

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