Preparation method of freeze-dried pellets for quantitatively preserving microorganisms

A vacuum freeze-drying and microbial technology, which is applied in the preservation of microorganisms, etc., can solve the problems of uncertainty in the quantitative results of microbial viable bacteria, and achieve the effects of avoiding sampling loss, good stability, and high survival rate of bacteria

Inactive Publication Date: 2017-07-07
NAT INST OF METROLOGY CHINA
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  • Abstract
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Problems solved by technology

[0002] Due to the quality of the medium, culture conditions, mixing degree of the bacterial solution, loss of microbial adsorption, volume of absorbed liquid, ac

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  • Preparation method of freeze-dried pellets for quantitatively preserving microorganisms

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[0029] The preparation method that the embodiment of the present invention provides comprises the following steps:

[0030] S101, after the target strain is inoculated on nutrient agar and enriched in single colony nutrient broth, the microbial bacterial liquid in the late logarithmic growth phase is obtained.

[0031] S102, diluting the microbial bacterial liquid, and using ultraviolet light absorption to assist in controlling the bacterial concentration to obtain a uniform bacterial suspension with a quantitative concentration.

[0032] Specifically, the microbial bacterial liquid is properly diluted, and ultraviolet light absorption (OD600) is used to assist in controlling the bacterial concentration to obtain a uniform bacterial suspension with a quantitative concentration.

[0033] S103, mixing the protective agent with sterile water to prepare a protective agent solution.

[0034] Specifically, the components of the protective agent may include protein, water-soluble su...

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Abstract

The embodiment of the invention relates to a preparation method of freeze-dried pellets for quantitatively preserving microorganisms. The preparation method comprises the following steps: inoculating target strains through nutrient agar and increasing bacteria of the target strains through single-colony nutrient broth, then obtaining a microorganism bacterium liquid of a later period of a logarithmic growth period; diluting the microorganism bacterium liquid and controlling bacterium concentration through assistant of ultraviolet light absorption to obtain a uniform bacterium suspension with a quantitative concentration; mixing a protection agent with bacterium-free water to prepare a protection agent solution; mixing the bacterium suspension with the protection agent solution to obtain a mixed solution, wherein the bacterium concentration of the mixed solution is 102cfu/50microliters-106cfu/50microliters; taking a quantitative amount of the mixed solution, freezing the mixed solution into pellets by using liquid nitrogen and then rapidly pre-freezing, wherein the pre-freezing time is not shorter than 2 hours, and the pre-freezing temperature is between 80 DEG C below zero and 20 DEG C below zero; and freeze-drying the pre-frozen pellets in vacuum to obtain freeze-dried pellets.

Description

technical field [0001] The invention relates to the technical field of microorganism freezing, in particular to a method for preparing freeze-dried pellets for quantitatively preserving microorganisms. Background technique [0002] Due to the quality of the culture medium, culture conditions, degree of mixing of the bacterial solution, loss of microbial adsorption, volume of absorbed liquid, accuracy of colony counting, and differences in the operation of different personnel, the quantitative results of microbial viable bacteria have great uncertainty. [0003] Article 25 Food inspection institutions shall be equipped with the necessary instruments and equipment, sample pre Processing device and standard material (reference material) or standard bacteria (poison) species, etc. Article 26 The instruments and equipment (including software), standard materials (reference materials) or standard bacteria (virus) species used by food inspection agencies shall be managed by specia...

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Application Information

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IPC IPC(8): C12N1/04
CPCC12N1/04
Inventor 傅博强陈敏璠林婧张玲隋志伟王晶
Owner NAT INST OF METROLOGY CHINA
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