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Biology

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Polysome Fractionation and Analysis of Mammalian Translatomes on a Genome-wide Scale

68.1K Views

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10:56 min

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May 17th, 2014

DOI :

10.3791/51455-v

May 17th, 2014

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Valentina Gandin¹, Kristina Sikström², Tommy Alain³, Masahiro Morita³, Shannon McLaughlan¹, Ola Larsson², Ivan Topisirovic¹

¹Lady Davis Institute and Department of Oncology, McGill University, ²Department of Oncology-Pathology, Karolinska Institutet, ³Goodman Cancer Centre and Department of Biochemistry, McGill University

Summary

Ribosomes play a central role in protein synthesis. Polyribosome (polysome) fractionation by sucrose density gradient centrifugation allows direct determination of translation efficiencies of individual mRNAs on a genome-wide scale. In addition, this method can be used for biochemical analysis of ribosome- and polysome-associated factors such as chaperones and signaling molecules.

Explore More Videos

MRNA Translation

Sucrose Density Gradient Centrifugation

Anota Algorithm

Genome wide Analysis

Protein Folding

Post translational Modifications

Chapters in this video

0:05

Title

1:54

Preparation of Sucrose Gradient

3:38

Isolation and Sedimentation of Polysomes

6:59

Polysome Fractionation and RNA Extraction

9:22

Results: Analytics of Translational Activity in MCF7 Cells Using Polysome Fractionation

10:24

Conclusion

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