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Rapid identification method of Saccharomyces cerevisiae haploid

A technology of beer yeast and haploid, applied in the field of microbial identification, can solve the problems of low haploid isolation rate and survival rate, lack of identification methods, and degradation of sporulation ability, so as to achieve authentic and repeatable experimental results Good, the effect of reducing workload

Inactive Publication Date: 2012-10-17
JIANGNAN UNIV
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Problems solved by technology

However, because industrial beer yeast lives in a nutrient-rich environment for a long time, the sporulation ability is degraded, the haploid segregation rate and survival rate are low, and there is a lack of effective identification methods, so there are not many studies in this area.

Method used

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  • Rapid identification method of Saccharomyces cerevisiae haploid
  • Rapid identification method of Saccharomyces cerevisiae haploid
  • Rapid identification method of Saccharomyces cerevisiae haploid

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example 1

[0025] Four suspected haploid strains (H2, H5, H18) preserved in the laboratory, one industrial strain of S. China University Industrial Microorganism Resource and Information Center, CICIM-CU) is used as the feasibility and accuracy of the experimental strain verification method of the present invention.

[0026] (1) Dilute the five strains of bacteria and haploid standard bacteria and spread them on the YEPD plate. After culturing at 28°C for 2 days, use a toothpick to pick up about 1 / 2 of the colonies into a 1.5mL centrifuge tube, and use ddH 2 O was washed 3 times, and colony PCR was performed after collecting the bacteria.

[0027] (2) Add 2 μL of each of the three primers, 8 μL of dNTP, 10 μL of 10×buffer, 0.5 μL of rTaq enzyme, and ddH to each reaction system (100 μL). 2 O 75.5 μL, mixed properly and placed in TECHNE TC-5000 PCR instrument for PCR reaction, the conditions are: denaturation at 95°C for 5 min; denaturation at 94°C for 1 min, annealing at 50°C for 30 s, e...

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Abstract

The invention discloses a rapid identification method of Saccharomyces cerevisiae haploid, and belongs to the field of microbial identification. The invention adopts an experimental strategy combining colony PCR and flow cytometry, and has high efficiency, accuracy, and good reproducibility. Compared with a traditional haploid identification method (long time cultivation, staining microscopy), the method of the invention can achieve experimental results in a very short period of time, so as to greatly reduce the workload and the working time, and obtain more reliable experimental results. Moreover, the method of the invention can simultaneously identify mating type ( MATa / MAT alpha) of haploid strains, and can minimize influence of Saccharomyces cerevisiae haploid standard bacteria on the experimental results. In the invention, MAT PCR is not only an identification method for mating types of strains, but also much of a primary judgment means for ploidy identification, and can fast and conveniently exclude the interference of some strains.

Description

technical field [0001] The invention relates to a method for identifying beer yeast haploid, in particular to a method based on colony PCR and flow cytometry, and belongs to the field of microbial identification. Background technique [0002] Yeast is the soul of beer brewing and plays a vital role in the quality of beer. Industrial beer yeast includes two types: upper yeast (ale brewing yeast, Saccharomyces cerevisiae) and lower yeast (lager brewing yeast, Saccharomyces pastorianus). More than 90% of commercially available beer is fermented by the lower yeast, so research on industrial beer yeast Also mostly for the yeast below. [0003] The following yeast, also known as Karl's yeast, are heterozygous polyploids or aneuploids. Their complex ploidy brings many difficulties to yeast improvement and genetic research. Yeast haploid strains have only one set of chromosomes, and their genes are simple and clear. They are often used as model strains for genetics and breeding wo...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
Inventor 李崎许维娜王金晶李永仙刘春凤郑飞云
Owner JIANGNAN UNIV
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