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Small-scale Nuclear Extracts for Functional Assays of Gene-expression Machineries
JoVE Journal
Biology
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JoVE Journal Biology
Small-scale Nuclear Extracts for Functional Assays of Gene-expression Machineries
DOI:

08:05 min

June 27, 2012

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Chapters

  • 00:05Title
  • 01:05Growing and Harvesting HeLa Cells for Nuclear Extraction
  • 01:56Swell HeLa Cells in Hypotonic Buffer
  • 03:00Lyse Cells using a Dounce Homogenizer
  • 04:27Salt-extract the Nuclei
  • 05:15Concentrate and Dialyze the Nuclear Extract
  • 06:16Results: Small-scale Nuclear Extract Technique for Functional Assays of Gene Expression Machineries
  • 07:51Conclusion

Summary

Automatic Translation

A protocol for preparation of robust, small-scale HeLa nuclear extracts is described. This protocol is valuable for assays that require use of small populations of cells, such as cells treated with drugs or RNAi. The method should be applicable to a wide variety of gene expression assays and other cell types, including patient cells.

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