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Snelle in vivo fixatie en isolatie van translationele complexen uit eukaryote cellen
JoVE Journal
Biology
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JoVE Journal Biology
Rapid In Vivo Fixation and Isolation of Translational Complexes from Eukaryotic Cells
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Snelle in vivo fixatie en isolatie van translationele complexen uit eukaryote cellen

Nederlands

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3,473 Views

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14:29 min

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December 25, 2021

DOI:

10.3791/62639-v

DOI:
10.3791/62639-v

•

14:29 min
December 25, 2021

•

3 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

Summary

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We presenteren een techniek om translationele (eiwitbiosynthese) complexen snel te stabiliseren met formaldehyde crosslinking in levende gist- en zoogdiercellen. De aanpak maakt het mogelijk om transiënte tussenproducten en dynamische RNA:eiwitinteracties te ontleden. De crosslinked complexen kunnen worden gebruikt in meerdere downstream-toepassingen, zoals in op diepe sequencing gebaseerde profileringsmethoden, microscopie en massaspectrometrie.

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Cite this Article

Janapala, Y., Woodward, K., Lee, J., Rug, M., Preiss, T., Shirokikh, N. E. Rapid In Vivo Fixation and Isolation of Translational Complexes from Eukaryotic Cells. J. Vis. Exp. (178), e62639, doi:10.3791/62639 (2021).

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