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Method for detecting fungal diversity in traditional soybean paste fermentation process

A fermentation process and diverse technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of heavy workload and inaccurate detection results, and achieve high sensitivity, repeatability and reliability , The test process is simple and convenient

Inactive Publication Date: 2012-07-18
HEILONGJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The present invention aims to solve the problem of heavy workload and inaccurate detection results in the traditional method for detecting fungal diversity in the fermentation process of soybean paste, and provides a method for detecting the diversity of fungi in the fermentation process of traditional soybean paste

Method used

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  • Method for detecting fungal diversity in traditional soybean paste fermentation process
  • Method for detecting fungal diversity in traditional soybean paste fermentation process
  • Method for detecting fungal diversity in traditional soybean paste fermentation process

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specific Embodiment approach 1

[0009] Specific embodiment one: the method for this embodiment detection fungal diversity in traditional bean paste fermentation process, carry out according to the following steps: one, take by weighing 10g bean paste sample, join in the phosphate buffer saline solution of 50mL 0.1mol / L with pipette Pipette the liquid until suspended, then add glass beads, shake for 5min, then centrifuge at 200r / min for 5min, and collect the supernatant; 2. Wash the precipitate with 0.1mol / L phosphate buffer, then centrifuge at 200r / min for 5min , collect the supernatant, repeat this step 3 times, 3, combine the supernatant obtained in step 1 and step 2, centrifuge at 9000r / min for 12min, discard the supernatant, collect the precipitate X, and dissolve the precipitate X with 5mL 0.1mol / L The phosphate buffer solution was washed 3 times to obtain the washed cells, and 8 mL of 0.1mol / L phosphate buffer solution was added to the cleaned cells, blown to suspend with a pipette, oscillated evenly, a...

specific Embodiment approach 2

[0013] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is that in step 3, the method for washing the precipitate X with 5 mL of 0.1 mol / L phosphate buffer solution is specifically: adding 5 mL of 0.1 mol / L phosphate buffer to the precipitate X Phosphate buffered saline solution, then blown to suspension with a pipette, and centrifuged at 9000r / min for 10min. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0014] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is: the reaction system of the first PCR amplification in step five is a 50 μ L reaction system, which consists of the following components:

[0015]

[0016] The PCR amplification conditions were: 94°C pre-denaturation for 2 min, 94°C denaturation for 30 s, 52°C annealing for 30 s, 72°C extension for 45 s, a total of 35 cycles, 72°C extension for 7 min, and 4°C incubation. Others are the same as in the first or second embodiment.

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Abstract

The invention relates to a method for detecting fungal diversity in the traditional soybean paste fermentation process. The invention aims to solve the problems that the traditional method for detecting the fungal diversity in the soybean paste fermentation process is high in workload and detection results are inaccurate. The method comprises the following steps of: weighing a soybean paste sample, adding the soybean paste sample into a phosphate buffer solution, suspending, adding glass beads, oscillating, centrifuging, and collecting a supernatant; washing a precipitate, centrifuging, and collecting a supernatant; centrifuging, abandoning a supernatant, washing a precipitate to obtain thalli, adding the phosphate buffer solution into the thalli, blowing and beating until a suspension is obtained, and oscillating to obtain a pretreated sample; extracting deoxyribonucleic acid (DNA) of a fungal genome in the pretreated sample; performing polymerase chain reaction (PCR) amplification to obtain a product A; diluting the product A, and performing PCR amplification to obtain a product B; and loading the product B serving as a sample, and performing electrophoresis by using a denatured gradient gel electrophoresis device to finish the detection. The method is simple, and high in sensitivity, repeatability and reliability, and comprehensively reflects the diversity of flora structures.

Description

technical field [0001] The invention relates to a method for detecting fungal diversity in the fermentation process of traditional soybean paste. Background technique [0002] There are many types of fungi in the fermentation process of soybean paste. These fungi have a great impact on the fermentation of soybean paste, which can change the fermentation speed and product flavor of soybean paste, and even cause fermentation failure. Finding out the diversity of fungi in the fermentation process of soybean paste is of great significance for realizing the industrial production of soybean paste. However, in the existing research on the types and quantities of fungi in the fermentation process of soybean paste, most of them are based on the traditional isolation and cultivation method, that is, the results of morphological identification. The traditional method of counting viable bacteria is limited to cultivable microorganisms, and many microorganisms are not cultivable. , thus...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
Inventor 葛菁萍宋刚凌宏志赵丹高冬妮柴洋洋陈丽
Owner HEILONGJIANG UNIV
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