Preparation of the Density Gradient Fraction System and Sucrose Gradients
3:04
Preparation of Cell Extracts
5:10
Loading Extracts onto Sucrose Gradients and Ultracentrifugation
6:03
Fractionation of Cytoplasmic Extracts for Polysome Profiling
6:46
Extraction of Protein and RNA for Analysis
8:45
Results: Polysome Profiling Detects Changes in Translation Initiation in Response to Stress Conditions
10:01
Conclusion
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.
