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In vivo Interrogation of Central Nervous System Translatome by Polyribosome Fractionation
Journal JoVE
Neurosciences
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Journal JoVE Neurosciences
In vivo Interrogation of Central Nervous System Translatome by Polyribosome Fractionation

In vivo Interrogation of Central Nervous System Translatome by Polyribosome Fractionation

DOI:
10.3791/51255-v

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09:13 min

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April 30, 2014

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, , ,
1Department of Molecular Neurobiology,German Cancer Research Center (DKFZ)

Chapitres

  • 00:05Titre
  • 01:45Spinal Cord and Brain Tissue Preparation
  • 03:28Myelin Flotation
  • 04:33Sucrose Gradient Fractionation
  • 06:23RNA Isolation from Individual Fractions
  • 07:36RNA Profiles from Spinal Cord and Brain Tissue
  • 08:25Conclusion

Summary

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This protocol illustrates essential modifications of polyribosome fractionation in order to study the translatome of in vivo CNS samples. It allows global assessment of translation and transcription regulation through the isolation and comparison of total RNA to ribosome bound RNA fractions.

Tags

Keywords: Polyribosome FractionationTranslatomeGene ExpressionMRNA TranslationCentral Nervous SystemNeurobiologyTranscriptomicsProteomicsCycloheximideMyelinSucrose Gradient

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