JoVE Journal

Genetics

This content is Open Access.

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

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.

Chapters in this video

0:04

Title

0:53

Bacterial Cultivation and Extraction of Genomic DNA

1:54

Multiplex PCR Setup and Cycling Conditions

3:14

PCR Amplicon Confirmation by Gel Electrophoresis

4:37

Capillary Electrophoresis Setup and Run Conditions

6:06

VNTR (Variable-number Tandem-repeat) Size Calling, Repeat Count Calculation and MLVA (Multi-locus Variable-number Tandem-repeat Analysis) Profiling

7:36

Minimum Spanning Tree Cluster Analysis of MLVA Data

9:21

Results: Gel Electrophoresis, Electropherograms, and Minimum Spanning Tree

10:09

Conclusion

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