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

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

Article DOI: 10.3791/62639-v 14:29 min December 25th, 2021
December 25th, 2021

Kapitel

Summary

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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Keywords: In Vivo Fixation Translational Complexes Eukaryotic Cells Rapid Chilling Formaldehyde Fixation Glycine Quenching Cell Lysis Ribosomal Fractions Affinity Purification RNA Analysis Mass Spectrometry Yeast Culture Exponential Growth Phase Formaldehyde Concentration Fixation Time Cell Pelleting Glycine Buffer
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