LAMP (Loop-mediated Isothermal Amplification) detection kit and method of pathogenic aeromonas hydrophila
A technology of Aeromonas hydrophila and a detection kit, which is applied in the field of detection of target DNA fragments, can solve the problems of numerous detection steps of pathogenic Aeromonas hydrophila, difficult accuracy and time-consuming data processing, etc. Achieve the effect of reducing background influence, convenient method and enhancing specificity
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[0052] The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.
[0053] 1) DNA extraction
[0054] Pick suspected colonies and add them to 0.4mL TE buffer (pH8.0) for shaking and mixing. Add 100 μL of lysozyme solution, incubate at 37°C for 10 minutes, add 10 μL of proteinase K solution, incubate at 55°C for 1-2 hours, add the same volume of Tris saturated phenol (pH8.0), shake vigorously, centrifuge at 11000g for 3 minutes, and take the supernatant solution, and repeated phenol extraction; take the supernatant, add 0.1 times the volume of sodium acetate (3mol / L), mix well, add an equal volume of ice ethanol, mix well, let stand at low temperature for 30 minutes, and centrifuge at 15000g for 5 minutes. Discard the supernatant, wash once with 70% cold ethanol, centrifuge at 15,000 g for 5 minutes at room temperature, discard the supernatant, add 50 μL TE solution to obtain a genomic DNA solution to be teste...
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