Simple method for fast extracting human fecal bacterium DNA

Abstract

The invention discloses a simple method for fast extracting human fecal bacterium DNA. The simple method sequentially includes the following steps that hot-cold rupturing treatment is carried out on a human fecal sample to rupture cells; the DNA is captured through AMPure XP magnetic bead liquid; magnetic force adsorption of magnetic beads and a magnetic separation frame is used for removing impurities and purifying the DNA. According to the extracting method, as a physical method (liquid nitrogen freeze thawing) and magnetic bead purification are organically combined, one operator can extract about 100 samples at the same time within one hour, which has great significance in comprehensively and fast researching people group intestinal floras and related disease occurrence mechanisms.

Description

technical field [0001] The invention belongs to the fields of biology and medicine; in particular, it relates to a simple method for rapidly extracting bacterial DNA from human feces. Background technique [0002] The intestinal microbial flora is a huge and complex ecosystem, which accounts for about 80% of the total number of microecological bacteria in the body. Contains 100 trillion kinds of microorganisms, involving 1000 types of bacteria, the number of which is 10 times the number of human cells in the immune system. These bacteria help the body digest food, produce vitamins to prevent diseases caused by bacteria in food, and stimulate the body. As research progressed, scientists discovered that gut microbes also play a key role in many chronic diseases and conditions, such as inflammation and obesity. There should also be a close relationship between the intestinal microbial flora and the host’s genetic factors. Human genes will have an important impact on the intes...

AI Technical Summary

Problems solved by technology

The phenol / chloroform / isoamyl alcohol extraction used in the traditional method will lose a part of nucleic acid every time it is extracted, and the alcohol precipitation efficiency of low concentration nucleic acid is low

The CTAB method (adding guanidine thiocyanate method) uses a CTAB-containing lysate, which has a significant effect on removing polysaccharide components in the sample, but washing to remove CTAB is more difficult than removing other salt ions, and a small amount of CTAB residues will also affect enzymes. Activity has a great influence

The kit method avoids the harm of phenol and chloroform to the human body, and the washing efficiency of the centrifugal column type is higher than that of the traditional washing efficiency, and the purity of the nucleic acid purified by the column type is also high. However, incomplete protease digestion sometimes occurs during the operation. Column clogging

Moreover, due to the differences in the extraction mechanism and steps of the above methods, the amount and purity of DNA obtained vary, resulting in different application effects in subsequent experiments, such as PCR amplification, DNA sequencing, and restriction endonuclease analysis.

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