Extracting method of total DNA of microorganisms in intestines of earthworms

A technology of intestinal microorganisms and extraction methods, which is applied in the field of extraction of total DNA of intestinal microorganisms in earthworms, can solve the problems of low extraction efficiency, affecting the extraction efficiency of total microbial DNA, and inability to overcome the interference of earthworm intestinal tracts, so as to improve the extraction efficiency, Improve the extraction efficiency and extraction quality, and overcome the effect of interference

Inactive Publication Date: 2016-08-03
NANJING UNIV
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Problems solved by technology

However, earthworm intestines contain a large amount of sediment, which seriously affects the extraction efficiency of total microbial DNA.
At present, there are few studies on the method of extracting the total DNA of earthworm intestinal microorganism

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  • Extracting method of total DNA of microorganisms in intestines of earthworms

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Embodiment 1

[0021] The method for extracting total DNA of earthworm gut microbes of the present invention includes the following steps:

[0022] Step 1. Place a piece of experimental earthworm on clean filter paper, collect the fresh feces of the earthworm, and store it in freezing at 4℃;

[0023] Step 2. After returning the earthworm to its original growth environment for 12 hours, place it on clean filter paper again to collect its fresh feces; mix the feces extracted twice and store them at 4°C for freezing;

[0024] Step 3. Take 0.2g of fresh earthworm feces (feces that are frozen and stored for no more than 24 hours) and place them in a centrifuge tube, add 20μL proteinase K and 100μL buffer, and use a shaker to shake for 2 minutes to mix, mix well and then at 56℃ Heat in a water bath for 60 minutes to obtain liquid phase A;

[0025] Step 4. Use the soil DNA extraction kit ( SpinKitforSoil kit) to extract DNA to obtain the total DNA of earthworm intestinal microorganisms;

[0026] Step 5. Use a

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Abstract

The invention discloses a method for extracting the total DNA of earthworm intestinal microorganisms. On the one hand, the extraction method takes fresh earthworm excrement as a sample, and performs pretreatment on the sample; on the other hand, combines proteinase K (digestion) with a soil DNA extraction kit Therefore, the extraction method of the present invention solves the problem that the existing methods cannot overcome the silt interference, and the extraction efficiency of total DNA of earthworm intestinal microorganisms is low; the extraction method of the present invention greatly improves the extraction efficiency and efficiency of total DNA of earthworm intestinal microorganisms. The extraction quality, the total DNA bands detected by 1% agarose gel electrophoresis are bright and clear, which can more accurately and comprehensively reflect the status of microorganisms in the gut of earthworms, laying the foundation for subsequent research on the structure and function of microbial communities.

Description

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Claims

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Application Information

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Owner NANJING UNIV
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