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Separating Bacteria by Capsule Amount Using a Discontinuous Density Gradient
JoVE Journal
Genetics
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JoVE Journal Genetics
Separating Bacteria by Capsule Amount Using a Discontinuous Density Gradient

Separating Bacteria by Capsule Amount Using a Discontinuous Density Gradient

DOI:
10.3791/58679-v

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05:52 min

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January 07, 2019

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, , , ,
1Wellcome Sanger Institute, Wellcome Genome Campus, 2Department of Medicine,University of Cambridge

Chapters

  • 00:04Title
  • 00:48Preparation of Bacterial Strains of Mutant Libraries
  • 01:40Preparation of Gradient Dilutions and Mini-gradient Test
  • 02:39Preparation of Discontinuous Density Gradients
  • 03:39Adding Prepared Cells to Gradients and Separation by Centrifuge
  • 04:30Results: Density Gradient Centrifugation Used to Separate Bacterial Strains Based on Capsule Production
  • 05:10Conclusion

Summary

Automatic Translation

Automatic Translation

We demonstrate the use of discontinuous density gradients to separate bacterial populations based on capsule production. This method is used to compare capsule amount between cultures, isolate mutants with a specific capsule phenotype, or to identify capsule regulators. Described here is the optimization and running of the assay.

Tags

Keywords: – Bacteria Separation – Capsule Amount – Discontinuous Density Gradient – Klebsiella – Capsule Regulation – Visual Demonstration – Density Gradient Medium – Centrifugation – Cell Suspension – Gradient Dilutions – Gradient Interface

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