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Analysis of Translation Initiation During Stress Conditions by Polysome Profiling
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Analysis of Translation Initiation During Stress Conditions by Polysome Profiling

Analysis of Translation Initiation During Stress Conditions by Polysome Profiling

DOI:
10.3791/51164-v

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10:59 min

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May 19, 2014

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, ,
1Department of Molecular Biology, Medical Biochemistry, and Pathology, Faculty of Medicine,Laval University, 2CHU de Quebec Research Center

Chapters

  • 00:05Title
  • 01:45Preparation of the Density Gradient Fraction System and Sucrose Gradients
  • 03:04Preparation of Cell Extracts
  • 05:10Loading Extracts onto Sucrose Gradients and Ultracentrifugation
  • 06:03Fractionation of Cytoplasmic Extracts for Polysome Profiling
  • 06:46Extraction of Protein and RNA for Analysis
  • 08:45Results: Polysome Profiling Detects Changes in Translation Initiation in Response to Stress Conditions
  • 10:01Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we describe a method to analyze changes in the initiation of mRNA translation of eukaryotic cells in response to stress conditions. This method is based on the velocity separation on sucrose gradients of translating ribosomes from non-translating ribosomes.

Tags

Polysome ProfilingTranslation InitiationStress ConditionsEukaryotic CellsMRNA TranslationDensity Gradient FractionationRibosome PopulationsTranslational ControlNeurodegenerative DiseasesCancer Development

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