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
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[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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