8. PFGE: Tips and Tricks to Success and Interpretation of Results for ...

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PulseNet Standardized Protocol for PFGE patient specimen collection pure culture grow isolated colony = + cell lysis and plug washing cells trapped in plug agarose cell suspension restriction electrophoresis imaging tiff for analysis
Interpretation of PFGE Data • Technical artifact vs. genetic variation – Technical artifact • Effect of reproducibility • “Ghost” or “shadow” bands • Effect of resolution • Pattern analysis – Genetic variation • Expected degree of variability • Dependent on the organism being studied
Page 1: PFGE: Tips and Tricks to Success an
Page 5 and 6: Reproducibility 3 isolates of Salmo
Page 7 and 8: Artifacts • Salmonella Saintpaul
Page 9 and 10: “Ghost” or “Shadow” Bands
Page 11 and 12: Washing PFGE plugs • Washes - was
Page 13 and 14: Incomplete cell lysis Undigested, g
Page 15 and 16: High background • DNA degraded
Page 17 and 18: Enzyme digestion without BSA BSA BS
Page 19 and 20: Star activity looks like incomplete
Page 21 and 22: Impact of run length on resolution
Page 23 and 24: Interpretation of PFGE Data • Tec
Page 25 and 26: Addition of a restriction site x x
Page 27 and 28: Mode of Transmission Contrasting Sh
Page 29: Acknowledgements All PulseNet parti

8. PFGE: Tips and Tricks to Success and Interpretation of Results for ...

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