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.

Inactive Publication Date: 2010-08-25
黑龙江省乳品工业技术开发中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide a method for identifying Lactobacillus plantarum in order to solve the problems of long period and high cost of the existing identification method of Lactobacillus plantarum

Method used

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  • Method for identifying plant lactobacillus
  • Method for identifying plant lactobacillus
  • Method for identifying plant lactobacillus

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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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Abstract

The invention discloses a method for identifying plant lactobacillus, which relates to an identifying method. The method solves the problems that the existing method for identifying the plant lactobacillus has long period and high cost. The method comprises the following steps: 1, performing PCR amplification by using a primer containing 22 basic groups; and 2, performing electrophoresis detection so as to realize identification of the plant lactobacillus. The identifying method of the invention has short period, and finishes the identification by only about 2 hours; and compared with the existing identifying method, the identifying method of the invention can identify the plant lactobacillus without special complex instruments and data processing, and has low identifying cost.

Description

technical field [0001] The invention relates to an identification method. Background technique [0002] Lactobacillus plantarum ( L. plantarum ) is a probiotic of the genus Lactobacillus. Lactobacillus plantarum can produce various natural antibacterial substances such as organic acids, bacteriocins, hydrogen peroxide, and diacetyl. It can maintain the balance of intestinal flora, improve the body's immunity, and promote nutrition. Many functions such as substance absorption, especially the antibacterial active metabolite bacteriocin produced by Lactobacillus plantarum, can not only improve intestinal ecology, but also be used for food and feed preservation. Lactobacillus plantarum is usually isolated from dairy products, because there are many types of microorganisms in dairy products (mainly including Lactobacillus plantarum, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus delbrueckii, Lactococcus lactis, Leucobacterium enterica Bacteria and Streptococcus t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
Inventor 姜毓君满朝新诸晓强迟涛胡博韬房玉国王旻秦兰霞周艳秋韩琳琳胡惠秩
Owner 黑龙江省乳品工业技术开发中心
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