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Rapid In Vivo Fixation and Isolation of Translational Complexes from Eukaryotic Cells
JoVE Journal
Biologia
Author Produced
This content is Free Access.
JoVE Journal Biologia
Rapid In Vivo Fixation and Isolation of Translational Complexes from Eukaryotic Cells

Rapid In Vivo Fixation and Isolation of Translational Complexes from Eukaryotic Cells

DOI:

14:29 min

December 25, 2021

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Capitoli

  • 00:10Introduction
  • 01:06Yeast Cell Culture and FIxation
  • 03:58Yeast Cell Disruption and Cytosol Collection
  • 05:45Mammalian Cell Culture and Fixation
  • 09:40Mammalian Cell Disruption and Cytosol Collection
  • 11:02Risultati
  • 13:41Conclusion

Summary

Traduzione automatica

We present a technique to rapidly stabilize translational (protein biosynthesis) complexes with formaldehyde crosslinking in live yeast and mammalian cells. The approach enables dissecting transient intermediates and dynamic RNA:protein interactions. The crosslinked complexes can be used in multiple downstream applications such as in deep sequencing-based profiling methods, microscopy, and mass-spectrometry.

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