Method for identifying plant lactobacillus
A Lactobacillus plantarum and product technology, applied in the field of identification, can solve the problems of high cost and long identification method cycle, and achieve the effects of short cycle, easy analysis, and low identification cost.
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specific Embodiment approach 1
[0010] Specific embodiment one: the method for identifying Lactobacillus plantarum in this embodiment is carried out according to the following steps: one, the sample to be identified uses DNA extraction kit to extract the DNA that obtains, carry out PCR amplification, the primer sequence of PCR amplification is 5'- CTACGGCAAGGCGACGCTGACG-3', 25 μl of PCR reaction system is composed of 2.5 μl of 10×PCRBuffer, 2.5 μl of MgCl with a concentration of 25 mmol / L 2 , 2.5μl of dNTPs with a concentration of 10mmol / L, 1.8μl of primers with a concentration of 10pmol / L, 2U of Taq polymerase, 1.0μl of DNA with a concentration of 50ng / ul and the rest of distilled water. The PCR reaction program is: 95°C Pre-denaturation for 5 minutes, 30 cycles, denaturation at 94°C for 30 seconds, annealing at 59°C for 30 seconds, extension at 72°C for 1 minute, and final extension at 72°C for 5 minutes; The sample to be identified with only one band in electrophoresis detection is Lactobacillus plantarum...
specific Embodiment approach 2
[0013] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is that the electrophoresis detection method in step 2 is: after mixing 10 μL of PCR product and 1 μL sample loading buffer, 1% agarose gel is used for electrophoresis detection, Electrophoresis was carried out for 30min at a constant voltage of 100V, and then the results were observed under ultraviolet light after EB staining. Other steps and parameters are the same as those in Embodiment 1.
specific Embodiment approach 3
[0014] Specific embodiment three: the method for identifying Lactobacillus plantarum in this embodiment is carried out according to the following steps: 1. The sample to be identified uses the DNA extracted by the DNA extraction kit to perform PCR amplification, and the primer sequence for PCR amplification is 5'- CTACGGCAAGGCGACGCTGACG-3', 25 μl of PCR reaction system is composed of 2.5 μl of 10×PCRBuffer, 2.5 μl of MgCl with a concentration of 25 mmol / L 2 , 2.5μl of dNTPs with a concentration of 10mmol / L, 1.8μl of primers with a concentration of 10pmol / L, 2U of Taq polymerase, 1.0μl of DNA with a concentration of 50ng / ul and the rest of distilled water. The PCR reaction program is: 95°C Pre-denaturation for 5 minutes, 30 cycles, denaturation at 94°C for 30 seconds, annealing at 59°C for 30 seconds, extension at 72°C for 1 minute, and final extension at 72°C for 5 minutes; If there is only one band in the electrophoresis test, it can be confirmed that the sample to be identif...
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