Kit and method for extracting pectin-rich polysaccharide plant DNA

A kind of polysaccharide and kit technology, applied in the field of genetic engineering, can solve the problems of difficult to obtain high-quality and high-quantity genomic DNA, inability to obtain DNA, and low DNA quantity, so as to improve DNA quality and yield and avoid secondary quality browning high-quality, high-quality

Active Publication Date: 2018-12-07
四川省植物工程研究院
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When using conventional kits, a large amount of polysaccharides will block the adsorption column, and the amount of extracted DNA is extremely low, or even DNA cannot be obtained
Therefore, it is difficult to obtain high-quality and high-quantity genomic DNA from plants rich in pectin polysaccharides using existing technologies

Method used

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  • Kit and method for extracting pectin-rich polysaccharide plant DNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Okra is one of the plants known to contain particularly high levels of pectin polysaccharides and polyphenols. Due to the combination of traditional buffers and viscous polysaccharides, it is easy to form insoluble jelly, which interferes with the extraction and purification of DNA. The present invention aims at this problem, improves the extraction method in combination with the kit described in the present invention, and successfully extracts high-quality okra genomic DNA. Specific steps are as follows:

[0057] (1) Take about 0.2 g of fresh young okra leaves in a mortar, add liquid nitrogen to grind them into fine powder, and quickly collect them into a 2.0 mL centrifuge tube.

[0058] (2) Add 1.5 mL of 4°C pre-cooled desugar buffer and 15 μL of β-mercaptoethanol to the centrifuge tube, mix well, and place in a 4°C refrigerator for 10 min, during which time, invert and mix once. Centrifuge at 9000 rpm for 8 min, discard the supernatant, and collect the precipitate. ...

Embodiment 2

[0069] Citrus, aloe vera, dendrobium, fig, and grape are plants containing pectin polysaccharides or other secondary metabolites that are particularly high. Using the method of the present invention to extract genomic DNA, the steps are as follows:

[0070] (1) Sample processing:

[0071] a. Citrus, figs, grapes: take the young leaves, add liquid nitrogen to grind them for later use;

[0072] b. Aloe vera: After removing the thorns, add liquid nitrogen to grind for later use;

[0073] c. Dendrobium: take the stem, add liquid nitrogen and grind for later use;

[0074] Take 0.2 g of the above-mentioned plant samples of a-c respectively, and put them into 2.0 mL centrifuge tubes for later use.

[0075] The rest of the steps were carried out in sequence with (2)-(11) in Example 1, and each plant DNA sample was collected.

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Abstract

The invention relates to a kit and a method for extracting pectin-rich polysaccharide plant DNA. The kit comprises a desaccharifying buffer solution, a lysate, leaching liquor, a column buffer solution, a protein-free solution, a rinsing solution, an eluant and a DNA adsorption column. An extraction method comprises the following steps: removing polysaccharide from a plant, extracting DNA, subsequently extracting and removing polysaccharide again by filtering with a column, thereby acquiring the extracted DNA with high quality and no impurity. The kit and the method provided by the invention are suitable for the extraction of plant genome DNA of mass secondary metabolites rich in polysaccharide, polyphenol, and the like. The kit has the characteristics of a low cost, simple, convenient andquick operation, high quality, high efficiency, high applicability, and the like.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a kit and a method for extracting plant DNA rich in pectin-like polysaccharides. Background technique [0002] In molecular biology experiments, the quality of DNA isolated from cells is crucial to many subsequent molecular biology experiments, for example, the success or failure of restriction enzyme analysis, Southern blot, DNA library construction, etc. Depends on the quality of the DNA, and the extraction method is an important factor affecting the quality of the DNA. At present, in the extraction of plant genomic DNA, the most commonly used methods are the classic CTAB (Cetyl-trimethyl-ammonium bromide, hexadecyltrimethylammonium bromide) method, SDS (Sodium dodecyl sulfate, dodecyl sulfate Sodium) methods, but these methods are generally only applicable to plants with low secondary substances such as polysaccharides and polyphenols. Due to the di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1006
Inventor 张景荣卓明刘玉珊叶昌华唐宵铧李臻杨马进王辉
Owner 四川省植物工程研究院
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