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A method for extracting genome dna from holly

A genome and holly technology, applied in the direction of recombinant DNA technology, DNA preparation, etc., can solve the problems of difficult to meet quality requirements, low concentration, low DNA quality, etc., achieve good DNA integrity, high DNA purity, and solve DNA yield low effect

Active Publication Date: 2019-05-03
SHENZHEN RES INST THE CHINESE UNIV OF HONG KONG
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  • Description
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Problems solved by technology

At present, the commonly used method is to extract the genomic DNA of Ilex chinensis, but the quality of the obtained DNA is not high and the concentration is low, which is difficult to meet the quality requirements of DNA library construction in high-throughput sequencing experiments.

Method used

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  • A method for extracting genome dna from holly
  • A method for extracting genome dna from holly
  • A method for extracting genome dna from holly

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Embodiment Construction

[0016] The present invention will be further described below with reference to the accompanying drawings and in combination with preferred embodiments.

[0017] The extraction of genomic DNA is a very important link in molecular biology experiments. The quality and yield of genomic DNA are of great significance for molecular biological research such as genomic DNA library construction. Phenolic substances and polysaccharides are two important factors affecting the extraction of plant genomic DNA, which often lead to the degradation of DNA molecules and seriously affect the purity and yield of DNA. The holly plant tissue is rich in sugars and phenols. Phenols are prone to oxidation and combine with DNA to form yellow or brown precipitates, which are difficult to dissolve. The purity of DNA is low and it is difficult to meet the needs of molecular biology research. Therefore, effective removal Polysaccharides, proteins, and phenolic substances are the key to extracting high-qual...

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Abstract

A method for extracting high-quality genome DNA from ammopiptanthus mongolicus includes the following steps of firstly, conducting liquid nitrogen quick freezing on plant materials, and grinding the plant materials into powder; secondly, adding a DNA extraction solution and protease K till the concentration of 20 micrograms per milliliter is reached, and keeping the temperature at 65 DEG C; thirdly, centrifugally collecting supernate, adding an equal-volume phenol:chloroform:isoamylol mixed liquid for extraction, and then centrifugally collecting supernate; fourthly, adding 0.6-0.8 fold volume of isopropanol to precipitate DNA, washing the DNA with ethyl alcohol, and partly drying the DNA and then dissolving the DNA in a TE buffer solution to obtain a DNA crude extract; fifthly, adding NaCl to the DNA crude extract till the concentration of 0.5 mol / L is reached, adding a CTAB / NaCl solution and then adding RNase A till the final concentration of 10 micrograms per milliliter is reached, and keeping the temperature at 65 DEG C; sixthly, sequentially conducting extraction through the equal-volume phenol:chloroform:isoamylol mixed liquid and equal-volume chloroform:isoamylol mixed liquid, and collecting supernate; seventhly, adding 0.6-0.8 fold volume of isopropanol to precipitate DNA, washing the DNA with ethyl alcohol, and partly drying the DNA and then dissolving the DNA in a TE buffer solution to obtain a high-quality DNA extraction solution. By means of the method, the high-quality genome DNA can be extracted from ammopiptanthus mongolicus.

Description

technical field [0001] The invention relates to plant DNA extraction, in particular to a method for extracting genome DNA from holly. Background technique [0002] Mongolian sand holly [Ammopiptanthus mongolicus (Maxim.) Cheng f.] is the subfamily of Fabaceae Papilionaceae. Sand holly is a super-xerophytic evergreen shrub. It is mainly distributed in the northwest (Xinjiang, Ningxia, Gansu) and Mongolia (the southeastern end of the Alxa Gobi region) in China. It is a national key protected plant. Sand holly has strong drought resistance, cold resistance and salt-alkali tolerance, and also has medicinal value. The research on the molecular mechanism of the stress resistance of Ilex chinensis has become a research hotspot in recent years. Ilex is a super-xerophytic shrub with high content of polysaccharides, phenols and other secondary metabolites. Previous studies have shown that the soluble sugar content of the leaves of Mongolian holly is up to 16%, and the polysaccharid...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
Inventor 何军贤冯磊
Owner SHENZHEN RES INST THE CHINESE UNIV OF HONG KONG
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