JoVE Journal

Chemistry

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Nanyang Technological University

Isolation and Preparation of Bacterial Cell Walls for Compositional Analysis by Ultra Performance Liquid Chromatography

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16.1K Views

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11:18 min

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January 15th, 2014

DOI :

10.3791/51183-v

January 15th, 2014

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Samantha M. Desmarais¹, Felipe Cava², Miguel A. de Pedro³, Kerwyn Casey Huang¹^,⁴

¹Department of Bioengineering, Stanford University, ²Department of Molecular Biology and Laboratory for Molecular Infection Medicine Sweden, Umeå Centre for Microbial Research, Umeå University, ³Campus de Cantoblanco, Universidad Autonoma de Madrid, ⁴Department of Microbiology and Immunology, Stanford University School of Medicine

Podsumowanie

The bacterial cell wall is composed of peptidoglycan, a macromolecular network of sugar strands crosslinked by peptides. Ultra Performance Liquid Chromatography provides high resolution and throughput for novel discoveries of peptidoglycan composition. We present a procedure for the isolation of cell walls (sacculi) and their subsequent preparation for analysis via UPLC.

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Bacterial Cell Wall

Cell Wall Composition

Cell Wall Isolation

High throughput Analysis

Rozdziały w tym wideo

0:05

Title

1:43

Day 1 - Lysing Bacterial Cultures

3:24

Day 2 - Enzymatic Digestions

6:28

Day 3 - Preparation of Samples for UPLC

9:29

Results: UPLC Chromatograms of PG Digested from E. coli Sacculi

10:36

Conclusion

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