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Single Particle Electron Microscopy Reconstruction of the Exosome Complex Using the Random Conical Tilt Method
JoVE Journal
Biology
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JoVE Journal Biology
Single Particle Electron Microscopy Reconstruction of the Exosome Complex Using the Random Conical Tilt Method
DOI:

12:10 min

March 28, 2011

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Chapters

  • 00:05Title
  • 01:11Principle of the Random Conical Tilt Method
  • 02:10Preparation of Holey Carbon Grids Covered with Thin Carbon
  • 05:08Negative Staining of the Exosome Complex
  • 06:23Electron Microscopy of the Exosome Complex
  • 08:10Image Processing of the Data
  • 10:55Representative Results for Single Particle Electron Microscopy Reconstruction of the Exosome Complex Using the Random Conical Tilt Method
  • 11:29Conclusion

Summary

Automatic Translation

This article describes a standard method to get a three-dimensional (3D) reconstruction of biological macromolecules using negative staining electron microscopy (EM). In this protocol, we explain how to get the 3D structure of the Saccharomyces cerevisiae exosome complex at medium resolution using the random conical tilt reconstruction method (RCT).

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