Method for extracting DNA from plant rich in polysaccharide and polyphenol

An extraction method and plant technology, applied in the field of molecular biology, can solve the problems of large loss of genomic DNA, reduced sequence length, incomplete structure, etc., and achieve the effects of inhibiting oxidation reaction, improving purity and concentration, and reducing activity

Active Publication Date: 2018-05-22
广州海思医疗科技有限公司
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  • Abstract
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Problems solved by technology

[0007] However, the current plant DNA extraction methods mostly focus on the removal of polysaccharides and polyphenols. However, the extracted genomic DNA has a large loss, incomplete structure, reduced sequence length, and low purity, which cannot meet the requirements of subsequent molecular analysis.

Method used

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  • Method for extracting DNA from plant rich in polysaccharide and polyphenol

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Experimental program
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Effect test

Embodiment 1

[0043] Embodiment 1, the preparation of silicon dioxide nanomaterial film:

[0044] A: Add the cage-type mesoporous silica nanomaterial into deionized water and stir evenly, then add N-carboxymethyl chitosan and stir evenly, then stir at 70°C for 4 hours, the silica nanomaterial and N-carboxylate The weight ratio of methyl chitosan is 3:2, filter, and wash 2 times with deionized water, collect powder after freeze-drying, obtain the modified silicon oxide nanomaterial;

[0045] B dissolving N-carboxymethyl chitosan in deionized medium and stirring evenly to prepare a film-forming solution of N-carboxymethyl chitosan with a mass concentration of 5%;

[0046] C, adding the modified silicon oxide nanomaterial obtained in step A to the film-forming liquid obtained in step B, the solid-liquid ratio of the modified silicon oxide nano-material to the film-forming liquid is 1g:5ml, drying, and forming a film , that is.

Embodiment 2

[0047] Embodiment 2, a kind of DNA extraction method that is rich in polysaccharide polyphenol plant

[0048] S1 Take the leaves of the plants to be extracted and place them under liquid nitrogen to fully grind them into fine powders to obtain sample powders. Transfer 100mg of sample powders to a pre-cooled 1.5ml centrifuge tube, add 1ml of 3×CTAB preheated at 65°C for cracking solution, mix it upside down, add 5 μl of RNase with a concentration of 100 mg / ml, bathe in water at 65°C for 20 minutes, and centrifuge at 12,000 rpm for 10 minutes;

[0049] The preparation method of described 3×CTAB lysate is: with 1.5g, the CTAB that pH value is 8.0, the sodium chloride of 4.091g, the Tris-HCl solution of 5ml, 20ml, the ethylenediaminetetraacetic acid solution that pH value is 8.0 , 2g N-carboxymethyl chitosan, 1g puerarin were added to the volumetric flask, mixed evenly, and then added DEPC treated water to make the volume to 50ml;

[0050] S2 Add an equal volume of Tris phenol-ch...

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Abstract

The invention belongs to the technical field of molecular biology and particularly relates to a method for extracting DNA from a plant rich in polysaccharide and polyphenol. The provided method for extracting DNA from the plant rich in polysaccharide and polyphenol comprises the following steps: using a specific lysis solution to lyse a cell, extracting the DNA with Tris phenol-chloroform mixed solution, chloroform and a sodium perchlorate combination solution, and purifying the extracted DNA through a self-made adsorption column membrane, thereby obtaining the high-quality genomic DNA. The purity of the DNA extracted through the method for extracting DNA from the plant rich in polysaccharide and polyphenol is high, the impurity is low, the DNA is further prevented from being damaged by asuperoxide anion free radical or active oxygen, the completeness of the extracted genomic DNA is guaranteed, and the method is favorable for the scientific researches like construction of the gene library, PCR (Polymerase Chain Reaction) analysis, and Southern hybridization.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a method for extracting DNA from plants rich in polysaccharides and polyphenols. Background technique [0002] DNA is the basic genetic material and the carrier of genetic information. A certain amount and high-quality DNA samples are the basis for restriction enzyme digestion, gene cloning, molecular hybridization, genetic polymorphism analysis and genomics and other molecular biology research. With the rapid development of high-throughput sequencing technology and the substantial reduction in sequencing costs, molecular biology research such as genome sequencing has become an important hot topic. Extracting high-quality genomic DNA is the basis for obtaining accurate test results, and is the basic and critical step for subsequent molecular analysis. [0003] Different plants or even the same type of plant tissue materials have different external properti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 周煌凯钟诗龙邓美英程祖福孙勇姚啟聪徐毓璇张秋雪
Owner 广州海思医疗科技有限公司
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