Method for rapidly extracting animal waste microbial genomes based on paramagnetic particle method

A technology of animal feces and microorganisms, applied in the biological field, can solve the problems of little understanding of microbial functions and metabolic mechanisms, incomplete microbial community structure and diversity, and achieve the effect of saving extraction time and simplifying experimental steps

Inactive Publication Date: 2015-10-28
FUJIAN NORMAL UNIV
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Problems solved by technology

For a long time, due to technical limitations, the understanding of the structure and diversity of microbial communities has not been comprehensive, and little is known about microbial functions and metabolic mechanisms.

Method used

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  • Method for rapidly extracting animal waste microbial genomes based on paramagnetic particle method

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Embodiment Construction

[0025] figure 1 Among them, lanes 1 and 2 are animal feces microbial genomes extracted by the method of the present invention, and lane 3 is DL4500Marker of TAKARA company.

[0026] A rapid extraction of animal feces microbial genome based on the magnetic bead method, the specific steps are as follows:

[0027] (1) Pretreatment of feces: Weigh two tubes of 0.3g fresh feces into 2ml centrifuge tubes, add 1ml Buffer A to shake for 1min, centrifuge at 3000rpm / min for 10min, transfer the supernatant to a new centrifuge tube, centrifuge at 10000rpm / min 10min. Discard the supernatant and save the pellet.

[0028] (2) Lysis of microbial cells in feces: Add 450 μl Buffer B and 50 μl Buffer C to the pellet, mix well and then bathe in water at 37°C for 1 hour, add 65 μl Buffer D and 65 μl Buffer E, mix well and bathe in water at 37°C for 2 hours.

[0029] (3) DNA dissociation and impurity protein extraction: add 600μl Buffer F to the solution, shake for 60s, centrifuge at 13000rpm / m...

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Abstract

The invention provides a method for rapidly extracting animal waste microbial genomes based on a paramagnetic particle method. According to the method for rapidly extracting the animal waste microbial genomes based on the paramagnetic particle method, microbial cells are broken through lysozyme, impurities are removed by use of a phenol chloroform isoamyl alcohol solution, and the silicon substrate magnetic bead is adopted to specifically adsorb the animal waste microbial genomes DNA. Compared with a conventional extracting method of the animal waste microbial genomes, the method for rapidly extracting the animal waste microbial genomes based on the paramagnetic particle method has the advantages that not only is a large amount of extracting time saved, but also experimental procedures are simplified, and the animal waste microbial genomes can be extracted more effectively, rapidly and economically.

Description

technical field [0001] The invention relates to a method for rapidly extracting animal feces microbial genome based on a magnetic bead method, and belongs to the field of biotechnology. technical background [0002] Microorganisms refer to the general term for a large number of extremely diverse, tiny biological groups that cannot be seen without a microscope. Therefore, microorganisms generally include viruses, subviruses (viroids, viroids, prions), eubacteria with prokaryotic cell structure, archaea, and fungi with eukaryotic cell structure (yeast, mold, mushroom, etc.), protozoa Animals and unicellular algae. [0003] The community structure and diversity of microorganisms in the environment, as well as the function and metabolic mechanism of microorganisms are research hotspots in microbial ecology. For a long time, due to technical limitations, the understanding of the structure and diversity of microbial communities has not been comprehensive, and little is known abo...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 王清水余彦
Owner FUJIAN NORMAL UNIV
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