Shigella sonnei special nucleotide PCR detection kit
A nucleotide and shigella technology, which is used in the determination/inspection of microorganisms, DNA/RNA fragments, biochemical equipment and methods, etc., can solve the problems of difficult identification, no rapid identification method, etc., and achieve high detection accuracy. , the market application prospects are wide, the effect of eliminating the extraction step
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[0036] 1: Genome extraction
[0037] (1) In the ultra-clean workbench, take out the strain from the strain preservation room, thaw it on ice, inoculate it into LB medium at a ratio of 1:1000, and cultivate overnight at 37°C on a shaker at 180rpm.
[0038] (2) Take 1 mL of the bacterial suspension into a 1.5 mL centrifuge tube, centrifuge at 8000 rpm for 5 minutes, and discard the supernatant.
[0039] (3) Add 250 μL of 50 mM Tris-HCl buffer (pH 8.0) to resuspend, centrifuge at 12,000 rpm for 5 minutes, discard the supernatant, and repeat once.
[0040] (4) Add 250 μL of 50 mM Tris-HCl buffer (pH 8.0) to resuspend, add 10 μL of 0.45M EDTA (pH 8.0), fully suspend, and incubate at 37°C for 20 minutes.
[0041] (5) Add 10 μL of 20 mg / mL lysozyme and incubate at 37°C for 20 minutes.
[0042] (6) Add 1.5 μL 20 mg / mL proteinase K and mix gently.
[0043] (7) Add 15 μL of 10% SDS, bathe in water at 50°C for 2 hours until the solution is clear, during which time gently invert and mi...
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