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

DOI :

10.3791/62639-v

14:29 min

December 25th, 2021

December 25th, 2021

3,157 Views

1Division of Genome Sciences and Cancer, The John Curtin School of Medical Research, The Australian National University, 2Centre for Advanced Microscopy, The Australian National University, 3Victor Chang Cardiac Research Institute

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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Rapid In Vivo Fixation

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