Rapid detection kit and detection method for multiple virulence factor GeXP of vibrio coralliilyticus
A Coral-lysing Vibrio and detection kit technology, applied in the biological field, can solve the problems of cumbersome process, easy to miss detection, unable to fully reflect the potential pathogenicity of pathogenic bacteria, etc.
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[0077] (1) Genomic DNA extraction of Vibrio corallilyticus
[0078] Firstly, the Vibrio coralliilyticus in the coral sample was purified and isolated on a plate, and then the Vibrio coralliilyticus was cultured.
[0079] ① Add 50 μL of 10% (w / v) sterile Chelex-100 solution into the centrifuge tube;
[0080] ② Pick a single colony from the culture plate of Vibrio coralliilyticus and put it into the above Chelex-100 solution;
[0081] ③ Place the centrifuge tube on a vortex mixer and shake it fully for 10 seconds, take a boiling water bath for 10 minutes, cool to room temperature, and shake it fully for another 10 seconds;
[0082] ④ Centrifuge at 13000r / min for 2min, and the supernatant is the genomic DNA of Vibrio corallilyticus, and store it in a -20°C refrigerator for later use.
[0083] (2) Design and synthesis of multigene PCR primers
[0084] Primers were designed for the virulence-related gene flhA, and the amplified fragment size was about 147bp:
[0085] flhA-F: AG...
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