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Absolute quantification probe and method for zygosaccharomyces bayer and application of probe and method

A technology of Zygosaccharomyces bayer and probes, which is applied to biochemical equipment and methods, measuring devices, determination/inspection of microorganisms, etc., can solve the problems of inappropriate timely detection, complicated operation of quantitative methods, and insufficient detection accuracy.

Active Publication Date: 2021-06-08
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] A kind of probe, kit and application for the absolute quantification of Zygosaccharomyces bailii of the present invention solve at least one of the following technical problems: (1) the existing method cannot realize the total amount detection of all Zygosaccharomyces bailii; (2) the existing Quantitative methods have low species resolution and / or insufficient detection accuracy; (3) Existing quantitative methods require high-cost instruments and / or strict operating environments, and are not suitable for timely detection after production sampling; (4) Existing quantitative methods are cumbersome to operate, etc.

Method used

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  • Absolute quantification probe and method for zygosaccharomyces bayer and application of probe and method
  • Absolute quantification probe and method for zygosaccharomyces bayer and application of probe and method
  • Absolute quantification probe and method for zygosaccharomyces bayer and application of probe and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1: Zygosaccharomyces Bailii Quantitative Probe Combination Reagent

[0060] Probe combination reagents; signal probe reagents and quenched probe reagents containing separate packages; wherein the signal probe sequences are shown in SEQ ID NO.2. .

[0061] Signal probe reagent and quenching probe reagent, is a dry powder or liquid shape; when dry powder, it can be diluted to a suitable concentration prior to experiment, for example, using sterile water or buffer to a concentration of 20 μm; for liquid However, the concentration can be 20-200 μm, and the reagent can be diluted before use, or it is used directly.

Embodiment 2

[0062] Example 2: Zygosaccharomyces Bailii Quantitative Kit and Its Use

[0063] Zygosaccharomyces Bailii Quantitative kit contains separately packaged signal probe reagents and quenched probe reagents; wherein the signal probe sequences are shown in SEQ ID NO.1, and the quenched probe sequence is shown in SEQ ID NO.2.

[0064] When the kit is used, it can be with heme, buffer, 2,2-ligand-bis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), H 2 O 2 With the use of.

[0065] How to use is:

[0066](1) Solution configuration. Configure 100 nm heme solution (reagent 1); TRIS-HCl having a final concentration of 50 mm, a final concentration of 50 mM, and finally pH of 7.9 (reagent 2); 7 mm 2,2-lienyl - double-( 3-ethylbenzene dihydrothiazoline-6-sulfonic acid) diammonium salt (ABTS) (reagent 3) and 7 mm H 2 O 2 Solution (reagent 4); the solvent is aseptic water.

[0067] (2) The signal probe forms a double strand with the sample DNA. 4 μl of the sample genomic DNA was a...

Embodiment 3

[0073] Example 3: Zygosaccharomyces Bailii Quantitative Kit

[0074] Zygosaccharomyces Bailii Quantitative kit contains separately packaged signal probe reagents and quenching probe reagents; wherein the signal probe sequences are shown in SEQ ID NO.1, and the quenched probe sequence is shown. .

[0075] The kit also contains 100 nm heme solution (reagent 1), Tris-HCl buffer, 7 mm 2,2-liazide-dual-(3-ethylbenzene dihydroxiazoline-6-sulfonic acid) ) Diammonium salt (ABTS), 7mm H 2 O 2 Solution.

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Abstract

The invention discloses a probe and a method for absolute quantification of zygosaccharomyces bayer and application of the probe and the method, and belongs to the fields of biology, fermentation and detection. According to the zygosaccharomyces bayer quantitative probe and the kit, total quantity detection of zygosaccharomyces bayer can be achieved, expensive instruments are not needed when the zygosaccharomyces bayer quantitative probe and the kit are used for detection and zygosaccharomyces bayer quantification, and quantification work can be rapidly completed within 2.5 h. Meanwhile, nucleic acid extraction does not need to be carried out on a sample used in the method. Based on the probe and the detection kit provided by the invention, the detection kit is used for quantifying the zygosaccharomyces bayer and has the characteristics of rapidness, convenience, cheapness and accuracy.

Description

Technical field [0001] The present invention relates to probes, methods and applications of Bayer bond yeast absolute quantities, belong to the field of biological fields, fermentation, detection. Background technique [0002] Zygosaccharomyces Bailii is a common yeast, such as red wine, tea, white wine, and vinegar in a variety of fermentation food systems. It plays an important role in the fermentation process, such as in wine research, Zygosaccharomyces Bailii and Saccharomyces Cerevisiae coincides with the content of ethyl acetate, and itself has strong flavor metabolism. It has been studied. Zygosaccharomycesbailii can produce 33 important volatile compounds, so track zygosaccharomyces bailii's biomass It is important to determine the stability of fermentation batch and the regulation of fermentation parameters. However, there are currently a lot of traditional fermentation food systems that are multi-strain co-fermentation system, which cannot judge the content of Zygosacch...

Claims

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Application Information

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IPC IPC(8): G01N21/64C12Q1/30
CPCG01N21/6428C12Q1/30G01N2021/6432
Inventor 吴群徐岩杜如冰
Owner JIANGNAN UNIV
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