A kind of preparation and use method of quality control strain quantitative pellet
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A technology of small balls and strains, which is applied in the field of preparation of quantitative small balls for quality control strains, can solve the problems of heavy workload and cumbersome operation, and achieve the effects of low cost, simple process and convenient use
Active Publication Date: 2022-07-12
青岛高科技工业园海博生物技术有限公司
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[0005] The invention provides a method for preparing and using quantitative pellets of quality control bacterial strains, which solves the technical problems of cumbersome operation and large workload in the prior art of bacterial liquid gradient dilution method
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preparation example Construction
[0022] A method for preparing quantitative pellets of quality control strains, comprising the following steps:
[0023] (1) The single colony of the quality control strain is picked into the liquid medium, cultivated to the logarithmic growth phase, and the bacterial liquid is obtained;
[0024] (2) take quantitative bacterial liquid to be diluted in freeze-dried bacterial strain protective agent to obtain protective agent bacterial liquid;
[0025] (3) take a quantitative protective agent bacterial liquid, and pre-freeze it after forming into a ball with liquid nitrogen;
[0026] (4) vacuum freeze-drying the pre-frozen pellets obtained in step (3) to obtain dry pellets;
[0027] (5) Packed into sterile vials, each sterile vial is equipped with a dry pellet.
[0028] In step (2), each 100ml of the freeze-dried bacterial strain protection agent includes the following components and their mass ratios: 10% protein, 5% water-soluble sugar, 10% freeze-dried plasticizer, and the b...
Embodiment
[0039] (1) The single colony of Escherichia coli of the quality control strain was picked into the tryptone soy peptone broth and cultivated to the logarithmic growth phase, that is, culturing at 36 degrees for 18-24 hours, to obtain a high-purity bacterial liquid with better activity;
[0040] (2) The freeze-dried strain protection agent includes the following components and their mass ratios: protein 10%, water-soluble sugar 5%, freeze-dried plasticizer 10%, the balance is water, and each 100ml freeze-dried strain protectant Including protein 10g, water-soluble sugar 5g, freeze-dried plasticizer 10g, and the balance is water. After all components are fully dissolved, autoclave at 115°C for 20 minutes, take it out, and cool it for later use. Among them, protein includes calf serum 3.0g and skim milk powder 7.0g, water-soluble sugar includes sucrose 3.0g and trehalose 2.0g; freeze-dried plasticizer includes polyethylene glycol 4000 3.0g, polyethylene glycol 6000 4.0g and Poly...
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Abstract
The invention relates to the technical field of preparation and use of microbial freeze-dried balls, in particular to a preparation and use method of a quality control strain quantitative ball. The preparation method includes the following steps: (1) picking a single colony of a quality control strain into a liquid medium, culturing it to a logarithmic growth phase, and obtaining a bacterial liquid; (2) taking a quantitative bacterial liquid into a freeze-dried bacterial strain protection agent, and using The protective agent is diluted to obtain a protective agent bacterial liquid; (3) a quantitative protective agent bacterial liquid is taken, frozen into spheres with liquid nitrogen, and then pre-frozen; (4) the pre-frozen pellets obtained in step (3) are subjected to vacuum freeze-drying to obtain a dry Small balls; (5) Packing. How to use: Add 1ml of sterile normal saline to the vial containing freeze-dried bacterial balls, dissolve and mix thoroughly, and then a bacterial liquid containing a certain number of bacteria can be obtained, which can be directly used in subsequent experiments. The quantitative pellet prepared by the invention has the beneficial effects of being able to meet the use requirements of quality control samples, simple in preparation method and convenient in use.
Description
technical field [0001] The invention relates to the technical field of preparation and use of microbial freeze-dried balls, in particular to a preparation and use method of a quality control strain quantitative ball. Background technique [0002] Laboratory data reliability and validity are the lifeblood of inspection work and scientific research. [0003] The use of microbial standard strains for testing methodology validation tests has received more and more attention. Following the requirements of the Chinese Pharmacopoeia for microbial testing methods applicability tests, relevant requirements have also been put forward in the risk monitoring of health food and cosmetics. The use of standard strains as reference standard substances to control the microbiological testing process of drugs, health foods and cosmetics is an important link to ensure scientific and accurate testing results. The "Chinese Pharmacopoeia", health food and cosmetics risk monitoring work manual, etc...
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