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Method for analyzing microbial community structure of fermented grain

A technology for microbial communities and fermented grains, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve problems such as incompatibility and difficult cultivation, and achieve the effect of saving time and simple operation.

Inactive Publication Date: 2013-04-10
LUZHOU PINCHUANG TECH CO LTD
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Problems solved by technology

The research on liquor microbes has a long history, but due to the limitations of technical methods, the understanding of the diversity of fermented grains microbial communities only depends on the classification and identification of traditional morphological characteristics, physiological and biochemical characteristics, and the traditional methods study the main components in the system. Cultivable microorganisms, suitable for the needs of difficult-to-cultivate and non-culturable microbial flora analysis

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  • Method for analyzing microbial community structure of fermented grain
  • Method for analyzing microbial community structure of fermented grain
  • Method for analyzing microbial community structure of fermented grain

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[0022] The technical scheme of the present invention is a method for analyzing the community structure of fermented wine, including extraction of genomic DNA, PCR amplification of 16S rDNA, DGGE electrophoresis (denaturing gradient gel electrophoresis), recovery of a single band of 16S rDNA after electrophoresis, Analyzing and recovering bands; the analyzing and recovering bands include the following steps: constructing a cloning library with 16S rDNA recovered after electrophoresis, enzyme digestion typing, sequencing and GenBank comparison analysis to obtain the community structure information of the microorganisms in the fermented wine.

[0023] Wherein, the genomic DNA extraction includes PBS (phosphate buffered saline) pretreatment and genomic DNA extraction of mashed wine samples. The PBS pretreatment includes the following steps: adding pH 7.4 PBS buffer and glass beads to the wine, vortexing, centrifuging at 200~500g (preferably 200g) for 5~10min, aspirating the supernatan...

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Abstract

The invention relates to a method for analyzing a microbial community structure of a fermented grain, belongs to the technical field of microbial ecology and aims to provide the method for analyzing the microbial community structure of the fermented grain. The technical scheme of the invention is as follows: the method for analyzing the microbial community structure of the fermented grain comprises extracting genomic DNA (Deoxyribose Nucleic Acid), expanding PCR (Polymerase Chain Reaction) of 16S rDNA, carrying out electrophoresis on DGGE (Denaturing Gradient Gel Electrophoresis), recycling a single strip of the 16S rDNA which is subjected to electrophoresis, and analyzing the recycled strip, wherein the step of analyzing the recycled strip comprises the following steps: establishing a clone bank by using the recycled 16S rDNA which is subjected to electrophoresis, carrying out enzyme segmentation, testing the sequence and carrying out comparison and analysis on GenBank so as to obtain the community structure information of the microorganism in the fermented grain. The method provided by the invention is widely applied to microbial community structure analysis and change regulation research in the white wine brewing process.

Description

Technical field [0001] The invention relates to a method for analyzing the microbial community structure of fermented wine, belonging to the technical field of microbial ecology. Background technique [0002] Denaturing gradient gel electrophoresis based on polymerase chain reaction (denaturing gradient gel electrophoresis) has been widely used in recent years as a molecular fingerprint analysis technique for microbial genetic diversity analysis in different environments. In 1993, Muyer et al. first applied this technology to the study of microbial ecology, and subsequently used it extensively in the study of genetic diversity and population differences in natural microbial communities. According to the melting characteristics of DNA, the denaturant concentration required for the denaturation of DNA double helices with different base composition is different. When mixed double-stranded DNA is electrophoresed on a polyacrylamide gel with a linear gradient of denaturant concentrati...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
Inventor 李德林王松涛许正宏敖宗华易彬
Owner LUZHOU PINCHUANG TECH CO LTD
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