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High-throughput microorganism isolation culture method

A technology for separation and cultivation of microorganisms, applied in the field of high-throughput separation and cultivation of microorganisms, which can solve the problems of increasing separation efficiency and "pollution" of dominant bacteria in the cultivation system, and achieve the effects of increasing separation efficiency, making full use of it, and improving utilization

Active Publication Date: 2016-09-21
OCEAN UNIV OF CHINA
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Problems solved by technology

[0006] The method of the present invention effectively solves the problem of "contamination" of the culture system by dominant bacteria in the single embedding method, ensures that the surface of the microspheres used for flow cytometry screening will not adhere to other microorganisms, and increases the separation efficiency

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  • High-throughput microorganism isolation culture method
  • High-throughput microorganism isolation culture method
  • High-throughput microorganism isolation culture method

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Embodiment 1

[0033] 1) Preparation of bacterial suspension

[0034] The samples of polymetallic nodules were collected from station PRZ1302-MC12 (154°49.8218′W, 10°29.2573′N) in the Eastern Pacific Nodule Area in 2013, with a water depth of 5098m. After the sampler obtains the sample, immediately put it into a sterile sealed bag and store it in the dark at 4°C. In the laboratory, take a piece of tuberculosis sample (diameter 1 ~ 2cm), put it into a 15ml chromatography column, add sterile filtered seawater (supplemented with 1mM NH 4 Cl), 10 ℃ dark enrichment for 6 months, every month with 7ml of fresh sterile seawater (supplemented with 1mM NH 4 C1) replace the culture medium. After the enrichment culture was completed, 1 ml of the culture solution was taken and filtered onto a 0.22 μm microporous membrane for microbial cell counting. According to the counting results, the concentration obtained by concentration of the sample is about 10 7 cells / ml bacterial suspension. Carry out doub...

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Abstract

The invention provides a high-throughput microorganism isolation culture method, which comprises the following steps: performing primary embedding on microorganisms to be isolated by taking agarose as an embedding material, performing secondary embedding to obtain microspheres by taking sodium alginate as an embedding material, and isolating the microspheres by virtue of a flow cytometer. According to the method, an in-situ sample can be directly added into a double-layer sphere culture system to implement co-culture of the microspheres and the in-situ sample. In such a co-culture manner, the difficult-to-culture microorganisms can be cultured more easily, and samples, particularly precious samples which are difficult to process such as nodules, can be fully utilized to increase the utilization rate of the precious samples.

Description

technical field [0001] The invention belongs to the technical field of microorganism separation and cultivation, and in particular relates to a high-throughput separation and cultivation method of microorganisms. Background technique [0002] With the rapid development of sampling technology, scientists have obtained more and more precious samples. How to efficiently isolate and culture microorganisms from these samples has become a hot issue for microbiologists in the world. Since the plate culture method was invented in 1887, this method has become a classic method for the isolation and culture of microorganisms. Then in 1979, scientists discovered that the strains obtained by the plate culture method were only a very small part of the total number. Since then, people have paid more and more attention to the shortcomings of the plate culture method. First, the proportion and quantity of nutrients in the plate are generally fixed. For samples from different sources, the p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/02C12N1/20C12N11/10C12N11/04
Inventor 张晓华张增虎武彦宏刘晨光肖天赵苑李诚博
Owner OCEAN UNIV OF CHINA
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