Method for extracting high-quality cell nucleus DNA (Deoxyribonucleic Acid) of plant rich in polysaccharide and polyphenol

Abstract

The invention discloses a method for extracting a high-quality cell nucleus DNA (Deoxyribonucleic Acid) of a plant rich in polysaccharide and polyphenol, relating to the field of plant functional genomics. The method comprises the following steps of: firstly, grinding a plant material by using liquid nitrogen; secondly, resuspending by using an HB (Haemoglobin) extraction buffer solution, mixing uniformly through volution and filtering; thirdly, centrifuging, washing precipitation with an extraction buffer solution 1 three times and resuspending; fourthly, carrying out water bath by using an extraction buffer solution 2 and RnaseH at the temperature of 65DEG C for 30 minutes; fifthly, extracting once by using chloroform-isoamylol and precipitating by using isopropanol; and sixthly, washing precipitation by using 75 percent pre-cooled ethanol 1-2 times and dissolving by using a TE (Tris-Ethylene Diamine Tetraacetic Acid) buffer solution. According to the pretreatment step in the methoddisclosed by the invention, DNAs of cell organelles such as mitochondria and chloroplast can be effectively removed; the inference on the nucleic acid extraction step caused by a large mount of secondary metabolites such as polysaccharide and polyphenol existing in the plant material is avoided; and the method is suitable for healthy tissues of wide plant species or frozen tissues with more complete cell structures and is especially suitable for high throughout sequencing in the subsequent researches.

Description

technical field [0001] The invention relates to the field of plant functional genomics, in particular to a method for extracting high-quality nuclear DNA of plants rich in polysaccharides and polyphenols; the method is fast and convenient, and is suitable for high-throughput sequencing or library construction. Background technique [0002] With the rapid development of high-throughput sequencing technology and the substantial reduction in sequencing costs, genome sequencing and resequencing have become an important part of plant genomics research. High-quality nuclear DNA (deoxyribonucleic acid) acquisition is the basis for high-throughput sequencing. At present, the nuclear DNA used for high-throughput sequencing or the construction of genomic libraries generally adopts the SDS method (sodium dodecylbenzenesulfonate), the CTAB method (cetyltrimethylammonium bromide) or the adsorption method based on silica gel matrix. Extraction by spin column and other methods. For plant...

AI Technical Summary

Problems solved by technology

In order to remove extranuclear genetic material such as chloroplast DNA, several existing genome extraction methods require yellowing treatment of plant materials, but the negative effect of yellowing treatment is that the growth state of the material becomes worse, and it often causes Induce secondary metabolites such as polyphenols to increase, thereby increasing the degree of polyphenol oxidation

And even after yellowing treatment, it is difficult to remove the pollution of mitochondrial DNA in higher plant cells

For tall perennial woody plants and most aquatic plants, it is difficult to obtain a large number of young and yellow leaves; The extraction method of high-quality nuclear DNA has become an urgent need for the genomics research of economic crops such as fruit trees and aquatic plants.

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