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Probe for absolute quantification of schizosaccharomyces pombe, kit and application

A technology of Schizosaccharomyces pombe and a detection kit, which is applied in the biological field and can solve the problems of insufficient detection accuracy, high cost of equipment for quantitative methods, and inapplicability of timely detection, etc.

Active Publication Date: 2021-05-11
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] A probe, kit and application for the absolute quantification of Schizosaccharomyces pombe of the present invention solve at least one of the following technical problems: (1) The existing method cannot realize the total amount of all Schizosaccharomyces pombe Detection; (2) 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 production after sampling (4) Existing quantitative methods are cumbersome to operate, etc.

Method used

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  • Probe for absolute quantification of schizosaccharomyces pombe, kit and application
  • Probe for absolute quantification of schizosaccharomyces pombe, kit and application
  • Probe for absolute quantification of schizosaccharomyces pombe, kit and application

Examples

Experimental program
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Effect test

Embodiment 1

[0059] Example 1: Schizosaccharomyces pombe quantitative probe combination reagent

[0060] Probe combination reagent; containing independently packaged signal probe reagent and quenching probe reagent; wherein, the signal probe sequence is as shown in SEQ ID NO.1, and the quenching probe sequence is as shown in SEQ ID NO.2 .

[0061] Signal probe reagents and quenching probe reagents are in dry powder or liquid form; in dry powder form, they can be diluted to an appropriate concentration before the experiment, for example, diluted with sterile water or buffer solution to a concentration of 20 μM; in liquid form , the concentration can be 20-200μM, and the reagent can be diluted before use, or used directly.

Embodiment 2

[0062] Embodiment 2: Schizosaccharomyces pombe quantitative kit and its use

[0063] The Schizosaccharomyces pombe quantitative kit contains independently packaged signal probe reagents and quenching probe reagents; wherein, the signal probe sequence is shown in SEQ ID NO.1, and the quenching probe sequence is shown in SEQ ID NO.2.

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

[0065] The method of use is:

[0066] (1) Solution configuration. Configure 100nM heme solution (reagent 1); configure Tris-HCL with a final concentration of 50mM, KCl with a final concentration of 50mM, and a final pH of 7.9 (reagent 2); 7mM 2,2-azino-bis-( 3-Ethylbenzodihydrothiazoline-6-sulfonic acid) diammonium salt (ABTS) (reagent 3) and 7mM H 2 o 2 Solution (reagent 4); solvents are sterile water.

[0067] (2) The signal probe forms a double strand with the sam...

Embodiment 3

[0073] Example 3: Schizosaccharomyces pombe Quantification Kit

[0074] Schizosaccharomyces pombe quantitative kit, containing independently packaged signal probe reagent and quenching probe reagent; wherein, the signal probe sequence is as shown in SEQ ID NO.1, and the quenching probe sequence is as shown in SEQ ID NO.2 .

[0075] The kit also contains 100nM heme solution (reagent 1), Tris-HCL buffer, 7mM 2,2-azino-bis-(3-ethylbenzodihydrothiazoline-6-sulfonic acid ) diammonium salt (ABTS), 7mM H 2 o 2 solution.

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Abstract

The invention discloses a probe for absolute quantification of schizosaccharomyces pombe, a kit and application, and belongs to the fields of biology, fermentation and detection. According to the probe for absolute quantification of schizosaccharomyces pombe and the kit, the total amount of schizosaccharomyces pombe can be detected, and can be used for detecting and quantifying the schizosaccharomyces pombe without expensive instruments, and the quantification work can be rapidly completed within 2.5h. Meanwhile, the sample used in the present invention need no nucleic acid extraction. Quantification of schizosaccharomyces pombe based on the probe and the detection kit provided by the invention has the characteristics of rapidness, convenience, cheapness and accuracy.

Description

technical field [0001] The invention relates to an absolute quantitative probe, kit and application of Schizosaccharomyces pombe, belonging to the fields of biology, fermentation and detection. Background technique [0002] Schizosaccharomyces pombe is a functional microorganism that widely exists in traditional brewing systems, such as liquor and wine. In the study of liquor, Schizosaccharomyces pombe can have strong acid resistance, and can reduce the accumulation of acetic acid and regulate the bacterial flora structure. Therefore, real-time tracking of the biomass of Schizosaccharomyces pombe is important for judging the stability of fermentation batches and the regulation of fermentation parameters. Guiding significance. However, at present, most traditional fermented food systems are multi-strain co-fermentation systems, and the content of Schizosaccharomyces pombe in samples cannot be judged by simple OD colorimetry, although fluorescent quantitative PCR combined wit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6816C12Q1/06C12N15/11C12R1/645
CPCC12Q1/6895C12Q1/6816C12Q2525/301C12Q2545/114C12Q2563/125
Inventor 吴群徐岩杜如冰
Owner JIANGNAN UNIV
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