Simple and fast extraction method for DNA in grape wine

Abstract

The invention discloses a simple and rapid DNA extraction method in wine, which comprises the following steps: A. 5g Silica is added with 5ml aqueous suspension for Silica precipitation to be evenly mixed after being cleaned by pure water and is set at the temperature of 4 DEG C for standby; B. wine with best adsorption conditions is then produced; C. the DNA of the wine is adsorbed and recycled: (1) Silica suspension is added to the wine mixture; (2) conservation is done for 1-2 hours under the temperature of 4DEG C, with constant shaking and oscillation; (3) centrifugation is done for 5 min under the temperature of 4DEG C with the amount of 5000g; (4) supernatant is removed and the Silica deposit is washed for 1-2 times by using washing fluid containing 0.1M Tris-HCl, pH 8.0, 1 percent of CTAB and 1M NaCl; (5) the centrifugation is done for 5 min with the amount of 5000g to collect the Silica deposit; (6) the Silica deposit is added with the pure water with the equivalent amount as the deposit and then well shaked and done with DNA heat elution after 5-10 min thermal insulation at the temperature of 65 DEG C; (7) centrifugation (10000g, 5min) is done to collect the supernatant containing DNA; (8) the supernatant in step (7) is extracted by chloroform with the same volume and the centrifugation (10000g, 3min) is done to collect top phase containing DNA; (9) the DNA in the supernatant is recycled and concentrated by using isopropanol precipitation; (10) the recycled wine DNA is then dissolved in 50 Mu lota TE (10mM Tris-HCl, pH 8.0) buffering agent.

Description

technical field [0001] The invention belongs to the technology of extracting DNA in wine, in particular to the DNA analysis technology for extracting wine DNA to identify the authenticity of wine. Background technique [0002] As we all know, wine is the most popular wine in the world, with high nutritional value and health care function. The founder of wine fermentation theory, the great French scientist Louis Pasteur (L. Pasteur) once said: "Wine is the healthiest drink". Medical research shows that wine contains about 600 kinds of ingredients beneficial to the human body, among which resveratrol (resveratrol) has the effect of preventing cancer and lowering cholesterol (Jang et al.1997). At present, more and more people drink wine, and famous and high-quality red (white) wine is favored by the world especially. In many developed countries, drinking wine has become a lifestyle. [0003] For a long time, the quality of various wines on the market has been uneven, with pr...

AI Technical Summary

Problems solved by technology

[0005] The main difficulties in extracting DNA from wine include: (1) the DNA content in wine is very small; (2) there are a large number of polyphenols (tannins), polysaccharides, organic acids, etc. in wine, which seriously interfere with DNA extraction and downstream utilization Molecular experiments of DNA (such as PCR); (3) The conventional isopropanol precipitation method is used for general wine operation, which is inconvenient; (4) Polyphenols and other pollutants are removed by dialysis before DNA extraction, or concentrated by freeze-drying and centrifugation Wine, etc. are time-consuming and require additional equipment; (5) Wine is weakly acidic, and its pH is generally between 3.4-4.2, which also limits the application of conventional DNA extraction methods