Detection method of germs in traditional zymotic soybean paste
A detection method and fermented soybean technology, which are applied in the directions of microorganism-based methods, biochemical equipment and methods, and microorganism determination/inspection, etc., can solve the problem of inability to accurately detect bacteria, etc. The effect of ensuring accuracy and reducing analytical bias
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specific Embodiment approach 1
[0023] Specific embodiment one: the detection method of bacteria in the traditional fermented soybean paste of the present embodiment is carried out according to the following steps: one, traditional fermented soybean paste sample bacterial total genome DNA extraction:
[0024] a. Pretreatment of traditional fermented soybean paste samples: Suspend 10 g of traditional fermented soybean paste samples with 50 mL of phosphate buffer solution with a pH value of 7.0 and a concentration of 0.1 mol / L, then add glass beads and vibrate for 5 minutes with a shaker, and then shake at 200 r Centrifuge for 5 min under the condition of 0.1 mol / L to collect the supernatant, then use the pH value of 7.0 and the concentration of 0.1 mol / L phosphate buffer to suspend, centrifuge and repeat 3 times, wherein each centrifugation speed is 200r / min , the centrifugation time is 5min, and then the centrifuged supernatant is collected and centrifuged at a speed of 9000r / min for 12min to collect the cent...
specific Embodiment approach 2
[0044] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is: in step 3, the sample volume is 5.0 μL each time, and the PCR amplification product obtained in 2.5 μL step 2 is mixed with 2.5 μL neutral loading solution composition. Other steps and parameters are the same as those in Embodiment 1.
[0045] In step 3 of this embodiment, the PCR amplification product obtained in step 2 is directly used without purification of the PCR amplification product.
specific Embodiment approach 3
[0046] Specific embodiment three: the difference between this embodiment and specific embodiment two is: 10 mL of neutral sample solution is composed of 2 g sucrose, 100 μ L of Tris-HCl with a concentration of 1 mol / L and a pH value of 7.8, and 20 μ L of Tris-HCl with a concentration of 0.5 mol / L L, EDTA with a pH value of 8.0, 10 mg of bromophenol blue and the remainder of deionized water. Other steps and parameters are the same as those in Embodiment 2.
[0047] In this embodiment, the neutral sample solution is stored in an environment of 4°C.
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