Quick extraction kit for common pathogenic bacterium genomes in food and method
A pathogenic bacteria and genome technology, which is applied in the rapid extraction kit and field of common pathogenic bacteria in food, can solve the problems of few applications and high price, and achieve the effect of wide application range, reduction of health damage and rapid extraction method.
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Embodiment 1
[0043] This example is to use the extraction method and kit of the present invention to test the extraction effect of 6 kinds of pathogen DNA simulated in different food samples.
[0044] 1. Make DNA extraction kits for 6 common pathogenic bacteria in food according to the following formula:
[0045] (a) Reagent A: PBS solution with pH 8.0;
[0046] (b) Reagent B: containing 70 mM SDS, 50 mM EDTA, 0.1 M Tris-Hcl, pH8.0;
[0047] (c) Reagent C: proteinase K at a concentration of 20 mg / mL;
[0048] (d) Reagent D: isopropanol;
[0049] (e) Reagent E: carboxylated magnetic bead solution;
[0050] (f) Reagent F: absolute ethanol;
[0051] (g) Reagent G: 70% ethanol;
[0052] (h) Reagent H: pH 8.0 TE solution containing 100 mM Tris-HCl (pH 8.0) and 10 mM EDTA.
[0053] 2. Listeria monocytogenes IQCC42222, IQCC42223; Staphylococcus aureus ATCC25923, ATCC27217; Salmonella IQCC10546, IQCC10547; Escherichia coli O157:H7 EDL933, IQCC10102; Cronobacter ATCC29544, ATCC51329; Shigella...
Embodiment 2
[0067] This example is to use the extraction method and kit of the present invention to extract 6 kinds of pathogenic bacteria DNA simulated in different food samples and compare them with the extraction effects of 2 commonly used commercially available kits, Wizard Genomic DNA Purification Kit and TIANampBacteria DNA Kit.
[0068] 1. Make DNA extraction kits for 6 common pathogenic bacteria in food according to the following formula:
[0069] (a) Reagent A: PBS solution with pH 8.0;
[0070] (b) Reagent B: containing 70 mM SDS, 50 mM EDTA, 0.1 M Tris-Hcl, pH8.0;
[0071] (c) Reagent C: proteinase K at a concentration of 20 mg / mL;
[0072] (d) Reagent D: isopropanol;
[0073] (e) Reagent E: carboxylated magnetic bead solution;
[0074] (f) Reagent F: absolute ethanol;
[0075] (g) Reagent G: 70% ethanol;
[0076] (h) Reagent H: pH 8.0 TE solution containing 100 mM Tris-HCl (pH 8.0) and 10 mM EDTA.
[0077] 2. Listeria monocytogenes IQCC42222; Staphylococcus aureus ATCC25...
Embodiment 3
[0092] This example is the detection result of real-time fluorescent PCR after using the extraction method and kit of the present invention to extract the DNA of 6 pathogenic bacteria in different food samples.
[0093] 1. Make DNA extraction kits for 6 common pathogenic bacteria in food according to the following formula:
[0094] (a) Reagent A: PBS solution with pH 8.0;
[0095] (b) Reagent B: containing 70 mM SDS, 50 mM EDTA, 0.1 M Tris-Hcl, pH8.0;
[0096] (c) Reagent C: proteinase K at a concentration of 20 mg / mL;
[0097] (d) Reagent D: isopropanol;
[0098] (e) Reagent E: carboxylated magnetic bead solution;
[0099] (f) Reagent F: absolute ethanol;
[0100] (g) Reagent G: 70% ethanol;
[0101] (h) Reagent H: pH 8.0 TE solution containing 100 mM Tris-HCl (pH 8.0) and 10 mM EDTA.
[0102] 2. Listeria monocytogenes IQCC42222, IQCC42223; Staphylococcus aureus ATCC25923, ATCC27217; Salmonella IQCC10546, IQCC10547; Escherichia coli O157:H7 EDL933, IQCC10102; Cronobacte...
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