Waters frequent food born pathogenic hacteria multiple PCR rapid detecting kit and its detecting method
A food-borne pathogenic bacteria and detection kit technology is applied in the field of Pseudomonas aeruginosa, Enterohemorrhagic Escherichia coli 0157 and Vibrio parahaemolyticus, and can solve the problems of many drug reagents, time-consuming and laborious, and high detection cost, Achieve the effect of short cycle, strong operability and low detection cost
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[0070] Example: multiplex PCR rapid detection kit for common foodborne pathogenic bacteria in water bodies
[0071] 1. Water sample collection and pretreatment
[0072] Collect 500ml of water samples in a sterile sampling bottle, centrifuge at 7000r / min for 20min, wash the precipitate with sterile distilled water three times, and then prepare DNA.
[0073] 2. DNA extraction
[0074] (1) Add 9.5mL TE to suspend the precipitate, add 0.5ml 10% SDS, 50μL 20mg / ml (or 1mg dry powder) proteinase K, mix well, and incubate at 37°C for 1 hour;
[0075] (2) Add 1.5ml 5mol / L NaCl and mix well;
[0076] (3) Add 1.5ml CTAB / NaCL solution, mix well, and keep warm at 65°C for 20 minutes; (4) Extract with an equal volume of phenol:chloroform:isoamyl alcohol (25:24:1), centrifuge at 5000r / min for 10 minutes, Transfer the supernatant to a clean centrifuge tube. ;
[0077] (5) Extract with an equal volume of chloroform:isoamyl alcohol (24:1), and transfer the supernatant to a clean tube;
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