DNA extraction method for polysaccharide-rich plant dried leaves

A technology of drying leaves and extraction methods, which is applied in the field of molecular biology, can solve the problems of difficult DNA extraction, achieve the effects of shortened extraction time, high purity, and simplified experimental procedures

Inactive Publication Date: 2014-04-30
SICHUAN AGRI UNIV
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  • Claims
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Problems solved by technology

[0005] The object of the present invention is to overcome the difficult problem of extracting DNA from the dry mature leaves of Prunus and other polysaccharide-rich plants, and provide a drying method suitable for polysaccharide plants. DNA Extraction Method of Leaves

Method used

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  • DNA extraction method for polysaccharide-rich plant dried leaves

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Embodiment Construction

[0035] The technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0036] A DNA extraction method suitable for dry leaves of polysaccharide plants, comprising the following steps:

[0037] 1) Take 0.5-1.0g each of Chinese cherry, European sweet cherry (red light), wild Chinese cherry and hairy cherry leaves dried on color-changing silica gel, add 0.3g of PVP and V C Add liquid nitrogen to the mortar and quickly grind it into powder. If the grinding is not enough, increase the number of grinding times with liquid nitrogen;

[0038] 2) Quickly transfer the ground leaf powder into 2.0ml centrifuge tubes, 3 tubes for each sample, add about 0.2-0.4g to each tube with 1.5ml 65°C preheated desugar buffer (1mol / L sodium chloride; 0.1 mol / L Tris-HCl PH8.0; 0.02mol / L EDTAPH8.0; 0.4mol / L glucose;) and 10μl β-mercaptoethanol, mix well and place in a water bath at 65°C for 10min;

[0039] 3) Ce...

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Abstract

The present invention discloses a DNA extraction method for polysaccharide plant dried leaves. The DNA extraction method comprises dried leaf grinding, desaccharifying, cracking, extraction, precipitation, washing, detection and other steps, wherein desaccharifying is the key step, a pre-heated desaccharifying buffer solution is added before the nuclear membrane is not cracked and the genome DNA is not released, the leaf tissue solution is suspended under a 65 DEG C water bath condition so as to dissolve polysaccharides and other secondary metabolites in the buffer solution, and low speed centrifugation is adopted to remove the polysaccharides and other impurities, such that the final DNA obtained through precipitation has high concentration and high purity. The DNA extraction method is suitable for dried mature leaves of all cerasus plants and other polysaccharide-rich plants, wherein the concentration of the obtained DNA can be increased if the leaves are the young and tender leaves, and the more DNA with high quality can be obtained from the mature leaves.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a method for extracting DNA from dried leaves of plants rich in polysaccharides, in particular to a method for extracting DNA from dried leaves of plants of the genus Sakura. Background technique [0002] Obtaining high-quality (high-concentration, high-purity, high-integrity) DNA is the most critical first step in molecular biology experiments. At present, a variety of methods have been developed to successfully extract DNA from plant leaves, callus, and tissue culture seedlings. , fruit, phloem and other tissues and organs to extract DNA. However, because most of the plant materials contain a wide variety of secondary metabolites, in some cases, different plants or even the same type of plant tissue materials have different sources, parts, external properties, and internal characteristics such as chemical components and tissue structures. Genomic DNA needs to choose a ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 王小蓉陈涛汤浩茹刘泽静陈清张勇罗娅黄晓姣陈娇
Owner SICHUAN AGRI UNIV
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