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Rapid In Vivo Fixation and Isolation of Translational Complexes from Eukaryotic Cells
JoVE Journal
Biyoloji
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JoVE Journal Biyoloji
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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Bölümler

  • 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
  • 11:02Sonuçlar
  • 13:41Conclusion

Özet

Otomatik Çeviri

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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