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Kit for rapidly extracting genomes of animal excrement by virtue of CTAB method

A technology of animal feces, genome, applied in the field of biology

Inactive Publication Date: 2015-10-14
FUJIAN NORMAL UNIV
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The study also found striking similarities among the gut microbiomes from the three geographically distinct populations, despite many differences

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  • Kit for rapidly extracting genomes of animal excrement by virtue of CTAB method

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Embodiment Construction

[0030] figure 1 Among them, lanes 1 and 2 are animal fecal microbial genomes extracted by this kit, and lane 3 is DL4500 Marker from TAKARA.

[0031] A kit for rapidly extracting animal feces genome based on the CTAB method, the specific steps are as follows:

[0032] (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.

[0033] (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. Add 100 μl Buffer F and 50 Buffer G solutions, and bathe in water at 65-80°C for 10-20min.

[0034] (3) DNA dissociation and impurity protein extraction: add 600μl Buffer H to the so...

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Abstract

The invention provides a kit for rapidly extracting genomes of animal excrement by virtue of a CTAB method. The kit is utilized for carrying out pretreating the excrement, splitting microbial cells in the excrement, dissociating DNA, abstracting impurity protein, adsorbing DNA by virtue of a DNA adsorption column, removing impurities and eluting DNA; after cells are disrupted, protein impurities are removed by virtue of a CTAB solution, meanwhile, genome DNA is adsorbed by virtue of a silicone mold adsorption column, and finally, the residual protein impurities are eluted by virtue of special deproteinization liquid and rinsing liquid. A conventional animal excrement microbial genome extraction method needs to be finished within 14 hours or longer, and the genomes are easily degraded due to the long-playing operation. Compared with the conventional animal excrement microbial genome extraction method, the kit has the advantages that the animal excrement genomes can be extracted within only 4 hours, so that plenty of extraction time is saved, meanwhile, experimental steps are simplified, and the microbial genomes of the animal excrement can be relatively effectively, rapidly and economically extracted.

Description

technical field [0001] The invention relates to a kit for rapidly extracting animal feces genome based on the CTAB method, which belongs to the field of biotechnology. technical background [0002] In the animal's body, there are countless tiny creatures that breathe and share fate with the animal all the time. These are the vast populations of microbes that live in the guts of animals. Studies have found that gut microbes are closely related to the immune system. In April 2012, Japanese scientists reported in "Science" that they found an immunosuppressive receptor that controls the composition of intestinal flora. If this receptor is missing, the micro-ecological environment in the intestinal tract will be disordered, which will lead to systemic An overactive immune system can lead to pathological changes such as autoimmune diseases. The researchers believe that these new findings may lead to the development of new ways to prevent or alleviate the symptoms of autoimmune ...

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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