Method for analyzing Bacillus community structure in white spirit fermentation system
What is Al technical title?
Al technical title is built by PatSnap Al team. It summarizes the technical point description of the patent document.
A technology of community structure and liquor, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc.
Inactive Publication Date: 2012-10-10
JIANGNAN UNIV
View PDF2 Cites 13 Cited by
Summary
Abstract
Description
Claims
Application Information
AI Technical Summary
This helps you quickly interpret patents by identifying the three key elements:
Problems solved by technology
Method used
Benefits of technology
Problems solved by technology
[0008] The present invention aims at the defects of the existing traditional microorganism separation method, and discloses a method using nested PCR combined with denaturing g
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more
Image
Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
Click on the blue label to locate the original text in one second.
Reading with bidirectional positioning of images and text.
Smart Image
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0038] p1-F: 5'-CGATGCGTAGCCGACCTGAG-3',
[0039] p1-R: 5'-AAGGAGGTGATCCAGCCGCA-3'.
[0040] The first-round PCR reaction system is 25 μL, including 12.5 μL 2×Taq PCR MasterMix enzyme, 0.5 μL (25 pmol) of each primer, 10.5 μL sterilized ultrapure water, and the amount of DNA template is about 10 ng. The first round of PCR reaction program was: 95°C pre-denaturation for 5 minutes, followed by 25 cycles, 95°C for 1 minute; 68°C for 90 seconds; 72°C for 2 minutes, and finally 72°C for 10 minutes. PCR products were detected by 1% agarose gel electrophoresis. The first-round PCR product was diluted 100-fold, and 1 μL was used as the template for the second-round PCR.
[0041] The primers used in the second round of PCR were:
[0044] The reaction system of the second round of PCR was the same as that of the first round. The second round of PCR reaction conditions wa...
Embodiment 2
[0048] p1-F: 5'-CGATGCGTAGCCGACCTGAG-3',
[0049] p1-R: 5'-AAGG AGGTGATCCAGCCGCA-3'.
[0050] The first-round PCR reaction system is 25 μL, including 12.5 μL 2×Taq PCR MasterMix enzyme, 0.5 μL (25 pmol) of each primer, 10.5 μL sterilized ultrapure water, and the amount of DNA template is about 10 ng. The first round of PCR reaction program was: 95°C pre-denaturation for 5 minutes, followed by 25 cycles, 95°C for 1 minute; 68°C for 90 seconds; 72°C for 2 minutes, and finally 72°C for 10 minutes. PCR products were detected by 1% agarose gel electrophoresis. The first-round PCR product was diluted 100-fold, and 1 μL was used as the template for the second-round PCR.
[0051] The primers used in the second round of PCR were:
[0054] The reaction system of the second round of PCR was the same as that of the first round.
[0055] The second round of PCR reaction conditio...
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more
PUM
Login to view more
Abstract
The invention discloses a method for analyzing the Bacillus community structure in a white spirit fermentation system, belonging to the technical field of microbial ecology. According to the invention, the diversity of a Bacillus community structure in the white spirit fermentation system is semi-quantitatively analyzed by a nested PCR-DGGE (polymerase chain reaction-denaturing gel gradient electrophoresis) method. The method comprises the following steps of: extracting the total genome DNA (deoxyribonucleic acid) in a yeast or fermented grains sample by a bead milling method; performing two steps of amplification of the Bacillus specific segment through the nested PCR; performing DGGE electrophoretic analysis; determining the microorganism species corresponding to the DNA stripe by use of a standard strain; performing glue cutting of the unknown stripe and recycling and then performing PCR again; connecting the T vector and cloning; performing DGGE electrophoresis again; performing positive cloning sequencing and performing blast comparison in the GenBank databae to obtain the microorganism information; digitally processing DGGE map by use of quantityone software; and calculating the proportion of different Bacillus strains in the total Bacillus flora according to the brightness of the stripe. The method disclosed by the invention has the advantages of simplicity, strong specificity and good repeatability, and lays foundation for further studying the importance of Bacillus in white spirit production.
Description
technical field [0001] A method for analyzing liquor fermentation system Bacillus The method of community structure, the present invention involves the use of modern molecular biology technology-polymerase chain reaction-deformation gradient gel electrophoresis (PCR-DGGE) to overcome the defects of traditional microbial isolation methods, qualitative and semi-quantitative analysis of microbial fermentation of Chinese liquor in the system Bacillus The structure and composition of the population and its dynamic evolution rules belong to the technical field of microbial ecology. Background technique [0002] The production of Chinese liquor is an open solid-state fermentation process. It can be said that the production of liquor actually includes a complex metabolic process of a huge microbial flora, which is greatly affected by the environment. As the environmental conditions change, the microflora will also change and affect the flavor and taste of liquor. Daqu and ferment...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more
Application Information
Patent Timeline
Application Date:The date an application was filed.
Publication Date:The date a patent or application was officially published.
First Publication Date:The earliest publication date of a patent with the same application number.
Issue Date:Publication date of the patent grant document.
PCT Entry Date:The Entry date of PCT National Phase.
Estimated Expiry Date:The statutory expiry date of a patent right according to the Patent Law, and it is the longest term of protection that the patent right can achieve without the termination of the patent right due to other reasons(Term extension factor has been taken into account ).
Invalid Date:Actual expiry date is based on effective date or publication date of legal transaction data of invalid patent.
Login to view more 
Login to view more