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Multi-locus Variable-number Tandem-repeat Analysis of the Fish-pathogenic Bacterium Yersinia ruckeri by Multiplex PCR and Capillary Electrophoresis
JoVE Journal
Genetics
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JoVE Journal Genetics
Multi-locus Variable-number Tandem-repeat Analysis of the Fish-pathogenic Bacterium Yersinia ruckeri by Multiplex PCR and Capillary Electrophoresis

Multi-locus Variable-number Tandem-repeat Analysis of the Fish-pathogenic Bacterium Yersinia ruckeri by Multiplex PCR and Capillary Electrophoresis

DOI:
10.3791/59455-v

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10:33 min

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June 17, 2019

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, ,
1Fish Health Research Group,Norwegian Veterinary Institute, 2University of Bergen

Chapters

  • 00:04Title
  • 00:53Bacterial Cultivation and Extraction of Genomic DNA
  • 01:54Multiplex PCR Setup and Cycling Conditions
  • 03:14PCR Amplicon Confirmation by Gel Electrophoresis
  • 04:37Capillary Electrophoresis Setup and Run Conditions
  • 06:06VNTR (Variable-number Tandem-repeat) Size Calling, Repeat Count Calculation and MLVA (Multi-locus Variable-number Tandem-repeat Analysis) Profiling
  • 07:36Minimum Spanning Tree Cluster Analysis of MLVA Data
  • 09:21Results: Gel Electrophoresis, Electropherograms, and Minimum Spanning Tree
  • 10:09Conclusion

Summary

Automatic Translation

Automatic Translation

The Multi-Locus Variable-number tandem-repeat Analysis (MLVA) assay presented here enables inexpensive, robust and portable high-resolution genotyping of the fish-pathogenic bacterium Yersinia ruckeri. Starting from pure cultures, the assay employs multiplex PCR and capillary electrophoresis to produce ten-loci MLVA profiles for downstream applications.

Tags

Keywords: Yersinia RuckeriMulti-locus Variable-number Tandem-repeat AnalysisMultiplex PCRCapillary ElectrophoresisGenotypingEpidemiologyPopulation StructureDiagnostic SpecificityInfection TracingVirulent ClonesEnvironmental Strains

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