Preparation and use methods of quality control strain quantitative pellet
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The technology of pellets and strains is applied in the field of preparation of quantitative pellets for quality control strains, which can solve the problems of large workload and complicated operation, and achieve the effects of low cost, simple process and reduced cell damage.
Active Publication Date: 2021-06-18
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 for quality control bacterial strains, comprising the steps of:
[0023] (1) pick a single bacterium colony of the quality control strain into the liquid medium, cultivate to the logarithmic growth phase, and obtain the bacterium liquid;
[0024] (2) Take the quantitative bacterial liquid and dilute it into the freeze-dried strain protective agent to obtain the protective agent bacterial liquid;
[0025] (3) Take the quantitative protective agent bacterial liquid, pre-freeze it after shaping it 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) Pack into sterile vials, and each sterile vial contains a dry pellet.
[0028] In step (2), each 100ml of the freeze-dried bacterial strain protection agent includes the following components and their mass proportions: 10% protein, 5% water-soluble sugar, 10% freeze-dried plasticizer, and ...
Embodiment
[0039] (1) Pick a single colony of Escherichia coli from the quality control strain and culture it in trypticase soytone broth to the logarithmic growth phase, that is, culture at 36 degrees for 18-24 hours to obtain a high-purity bacterial solution with better activity;
[0040] (2) The freeze-dried strain protectant includes the following components and their mass ratios: 10% protein, 5% water-soluble sugar, 10% freeze-dried plasticizer, and the balance is water. Every 100ml freeze-dried strain protectant It includes 10g of protein, 5g of water-soluble sugar, 10g of freeze-dried plasticizer, and the balance is water. After all components are fully dissolved, sterilize under high pressure at 115°C for 20 minutes, take it out, and cool it for later use. Among them, the protein includes calf serum 3.0g and skimmed milk powder 7.0g, water-soluble sugar includes 3.0g sucrose and trehalose 2.0g; freeze-dried plasticizer includes polyethylene glycol 4000 3.0g, polyethylene glycol 6...
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Abstract
The invention relates to the technical field of preparation and use of microbial freeze-dried pellets, in particular to preparation and use methods of a quality control strain quantitative pellet. The preparation method comprises the following steps: (1) picking a single colony of a quality control strain into a liquid culture medium, and culturing to a logarithmic phase to obtain a bacterial liquid; (2) adding a certain amount of the bacterial liquid into a freeze-dried strain protective agent, and diluting with the protective agent to obtain a protective agent bacterial liquid; (3) taking quantitative protective agent bacterial liquid, freezing the bacterial liquid into a pellet with liquid nitrogen, and pre-freezing the pellet; (4) performing vacuum freeze drying on the pre-frozen pellet obtained in the step (3) to obtain a dried pellet; and (5) sub-packaging. The use method comprises the following steps: adding 1 ml of sterile normal saline into a penicillin bottle filled with the freeze-dried pellets, fully dissolving and uniformly mixing to obtain a bacterial liquid containing a certain number of bacteria, and directly applying the bacterial liquid to subsequent experiments. The quantitative pellet has the beneficial effects that the use requirements of quality control samples can be met, the preparation method is simple, and the use is convenient.
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 quality control bacterial strain quantitative beads. Background technique [0002] The reliability and validity of laboratory data is the lifeline of inspection work and scientific research. [0003] The use of microbial standard strains for testing methodological verification tests has received more and more attention. Following the requirements of the "Chinese Pharmacopoeia" for the applicability test of microbial test methods, relevant requirements have also been put forward in the risk monitoring of health food and cosmetics. Using standard strains as reference standard substances to control the microbial testing process of drugs, health foods and cosmetics is an important link to ensure scientific and accurate test results. There are clear requirements in the "Chinese Pharmacopoeia", Health Food and ...
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