Method for extracting DNA of Chinese patent medicines or Chinese herbal medicine-containing health care products

An extraction method and technology of traditional Chinese medicinal materials, which are applied in the field of drug testing to achieve remarkable novelty and originality, and improve yield and quality.

Inactive Publication Date: 2015-10-28
MACAU UNIV OF SCI & TECH
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  • Description
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Problems solved by technology

[0004] Based on this, the purpose of the present invention is to provide an improved method for extracting the DNA of Chinese patent medicines or health products containing Chinese herbal medicines, aiming to solve the adverse effects caused by the addition of starch or drug-derived starch to the traditional CTAB method for DNA extraction. Thereby improving the extraction efficiency and quality of DNA

Method used

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  • Method for extracting DNA of Chinese patent medicines or Chinese herbal medicine-containing health care products

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Embodiment 1

[0020] A method for extracting the DNA of a Chinese patent medicine or a health product containing Chinese medicinal materials of the present embodiment, comprising the following steps:

[0021] (1) Put about 2 g of the sample fine powder shown in Table 1 into a 10 ml centrifuge tube, add 2.5 ml of 2×CTAB extraction buffer preheated at 65°C and 100 μL of mercaptoethanol, and lyse in a water bath at 65°C for 2 hours, 8 Centrifuge at 000rpm for 5min, during which time it is often inverted and mixed;

[0022] Table 1 The situation of ginseng medicine preparations used in the test

[0023]

[0024]

[0025] (2) Add 5ml of methanol to the supernatant, precipitate at -20°C for 1h, and centrifuge at 8000rpm for 5min;

[0026] (3) Transfer the precipitate to a 2ml EP tube, add 600 μL of 2×CTAB extraction buffer preheated at 65°C and 16 μL of mercaptoethanol, and lyse at 65°C for 20 minutes, shaking frequently during the period;

[0027] (4) Add an equal volume of chloroform: i...

Embodiment 2

[0036] A method for extracting the DNA of a Chinese patent medicine or a health product containing Chinese medicinal materials of the present embodiment, comprising the following steps:

[0037] (1) Take several fine powders of the samples listed in Table 1, put them in 2ml EP tubes, add 600 μl of 2×CTAB extraction buffer preheated at 65°C and 16 μl of mercaptoethanol, and lyse at 65°C for 2 hours, during which time Mix upside down;

[0038] (2) Add an equal volume of chloroform-isoamyl alcohol (24:1) and mix by inversion for 5 min, centrifuge at 12 000 rpm for 15 min; take the supernatant and add an equal volume of -20°C pre-cooled isopropanol, and precipitate at -20°C for 2 h. Centrifuge at 12 000rpm for 15min;

[0039] (3) Remove the solution in the lower layer and add sterile water to 600 μl, add 1200 μl of methanol and 60 μl of 3M sodium acetate solution, mix well, place at -20°C for precipitation for 1 hour, and centrifuge at 12 000 rpm for 15 minutes;

[0040] (4) Tak...

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Abstract

The invention discloses a method for extracting DNA of Chinese patent medicines or Chinese herbal medicine health care products, and a step of methanol precipitation of DNA is added in a traditional CTAB-method process. Through a cause that the DNA yield and quality are affected in a system screening CTAB-method extraction process, a fact that a starchy component in the Chinese patent medicines or Chinese herbal medicine health care products can form a high polar compound with CTAB so as to affect subsequent DNA isopropanol precipitation effect is found out for the first time; a fact that the step of methanol precipitation of DNA is added in any step in the traditional CTAB-method process and thus the yield and quality of DNA can be significantly improved and DNA molecular identification can be successfully carried out is found out.

Description

technical field [0001] The invention belongs to the technical field of drug detection, and more specifically, the invention relates to an optimized method for extracting DNA of Chinese patent medicines or health products containing Chinese medicinal materials. Background technique [0002] CTAB (hexadecyltrimethylammonium bromide, hexadecyltrimethylammonium bromide) is a cationic detergent that has the property of precipitating nucleic acids and acidic polysaccharides from solutions with low ionic strength. In solutions of high ionic strength (>0.7mol / L NaCl), CTAB forms complexes with proteins and polysaccharides, but cannot precipitate nucleic acids. The nucleic acid can be separated by organic solvent extraction to remove impurities such as proteins, polysaccharides, and phenols, and then add ethanol precipitation. [0003] During the traditional CTAB method for extracting DNA from Chinese patent medicines or health products containing Chinese medicinal materials, str...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 周华程春松刘良
Owner MACAU UNIV OF SCI & TECH
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