A method for rapid quantitative determination of microbial community composition during liquor fermentation

A microbial community and fermentation process technology, applied in the field of rapid PSP-qPCR quantification, can solve the problems of high cost, inability to meet the composition and structure of microbial community, and high quality requirements of endonucleases, so as to achieve low cost, improve wine production efficiency and effect significant effect

Active Publication Date: 2018-08-31
ANHUI RUISIWEIER TECH
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  • Claims
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Problems solved by technology

But the cost is expensive, the purity of DNA and the quality of endonuclease are high, etc.
[0011] However, none of the above-mentioned techniques can quickly determine the composition and structure of the microbial community in the liquor fermentation process in about two hours on site.

Method used

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  • A method for rapid quantitative determination of microbial community composition during liquor fermentation
  • A method for rapid quantitative determination of microbial community composition during liquor fermentation
  • A method for rapid quantitative determination of microbial community composition during liquor fermentation

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Embodiment Construction

[0043] In this example, the method for rapidly determining the composition of the microbial community in the liquor fermentation process is to use phylum-specific primers in the PCR reaction system to specifically amplify the microbial flora of a specific phylum during the liquor fermentation process, and use qPCR The system performs quantitative amplification to obtain quantitative identification.

[0044] Prepare gate-specific primers as follows:

[0045] Step a, extracting the metagenomic group of the brewing microbial community sample as a PCR amplification template;

[0046] Step b, amplifying the ribosomal DNA in the amplified template is close to the full-length 16s rDNA sequence, and the amplification primers are 27F (5'-AGA GTT TGA TCC TGG CTC AG-3') and 1492R (5'-GGT TAC CTT GTT ACG ACTT-3'), obtain the amplification product;

[0047] Step c. Perform TA cloning, blue-white screening and white-white colony bidirectional sequencing sequentially after the amplificatio...

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Abstract

The invention relates to a method for rapidly determining the microbial community composition in the liquor fermentation process, which is characterized in that the specific amplification of the microbial flora of a specific phylum is achieved in the liquor fermentation process by using door-specific primers in the PCR reaction system, And use the qPCR system for quantitative amplification to obtain quantitative identification. According to the known microbial phylum, the present invention designs primers for Bacteroidetes, Proteobacteria, Firmicutes, Syntrophyta and Actinomycetes respectively, and quantitative amplification based on phylum-specific primers can be performed at the phylum level Rapid investigation of the macroscopic structural changes of the microbial community during liquor fermentation.

Description

technical field [0001] The invention relates to a microbial community operation method in the field of biotechnology, and is a rapid PSP-qPCR quantitative method for determining the composition and structure of microbial communities in the liquor fermentation process based on phylum-specific primers (PSP). Background technique [0002] Natural complex microbial communities are generally composed of dozens, hundreds or even more different species of microorganisms, and the composition will change dynamically under different environmental conditions. Most of the bacterial species included in almost all natural microbial communities are poorly understood by humans, so it is still very difficult to quickly quantify the structural composition of natural microbial communities during liquor fermentation. Common techniques currently include: [0003] One is the traditional culture method; for example, most of the microorganisms in the pit mud belong to uncultivated bacteria, and th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/06C12R1/01
Inventor 何宏魁周庆伍李安军罗锡梅张会敏刘国英曹润洁汤知辉张治洲
Owner ANHUI RUISIWEIER TECH
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