High-efficiency extracting method of food-borne pathogen nucleic acid
A food-borne pathogenic bacteria and extraction method technology, applied in the field of efficient nucleic acid extraction of food-borne pathogenic bacteria, can solve the problems of tediousness, slow detection speed, time-consuming nucleic acid extraction and purification, etc., and achieve high purity and high yield Large, the effect of eliminating the interference of food and medium components
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Embodiment 1
[0024] A method for efficiently extracting nucleic acids from food-borne pathogenic bacteria, the composition of which includes: centrifuging the bacterial culture solution, adding buffer A to the centrifuged sediment, shaking and adding lysozyme, adding proteinase K after incubation, adding buffer B, shaking vigorously Afterwards, incubate at a temperature of 65°C for 10 minutes to make a lysis solution, pass through the silicon matrix SiO 2 The adsorption membrane of the material collects high-quality nucleic acid extraction nucleic acid solution.
Embodiment 2
[0026] In the method for efficiently extracting nucleic acid from food-borne pathogenic bacteria, the centrifugation of the bacterial culture solution is to centrifuge 1-5 mL of the bacterial culture solution at a speed of 8,000 rpm for 5 minutes, and suck up the supernatant.
Embodiment 3
[0028] The method for efficiently extracting nucleic acids from food-borne pathogenic bacteria includes adding buffer A to the centrifuge, shaking and adding lysozyme, adding proteinase K after incubation, adding buffer B, and shaking vigorously at temperature Incubate at 65°C for 10 minutes to prepare a lysis solution: add 250 μL of buffer A to the bacterial pellet: 20 mol / L TrisHCl, 2 mol / L EDTA, 1.2% polyethylene Diol octyl phenyl ether Tri ton, shake until the cells are thoroughly suspended, add lysozyme at a final concentration of 20mg / mL, incubate at 37°C for 30 minutes, add 20μL of protease at a concentration of 20mg / mL to the tube K solution, mix well, then add 200 μL buffer B: 0.05mol / L TrisHCl, 0.1mol / L sodium chloride NaCI with pH 7.6, 0.05mol / L EDTA, 2% Sodium Dodecyl Sulfate SDS, after vigorous shaking, incubate at 65°C for 10 minutes.
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