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Non-toxic extract solution combination gpr.1 for efficient extraction of plant genome dna and its extraction method

A plant genome and extract technology, applied in the biological field, can solve problems that have not been reported

Active Publication Date: 2021-09-14
TIANJIN AGRICULTURE COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the method and application of this type of extraction of genomic DNA have not been reported yet.

Method used

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  • Non-toxic extract solution combination gpr.1 for efficient extraction of plant genome dna and its extraction method
  • Non-toxic extract solution combination gpr.1 for efficient extraction of plant genome dna and its extraction method
  • Non-toxic extract solution combination gpr.1 for efficient extraction of plant genome dna and its extraction method

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Experimental program
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Embodiment 1

[0041] Preparation of the extract:

[0042] The preparation of the conventional SDS method extraction buffer will not be repeated, the following is the composition of GPR.1A, GPR.1D and GPR.1E solutions and their respective working concentrations.

[0043] Solution GPR.1A: 3.3% PEG8000, 0.5M NaCl, 0.1M Tris-HCl, 0.05M EDTA, 3.5% PVPP added before use, the balance is double distilled water;

[0044] Solution GPR.1D: 18.5% PEG8000, 3.5M NaCl, the balance is double distilled water;

[0045] Solution GNR.1E: 50mM Tris-HCl, 10mM EDTA, 2.5M NaCl, the balance is double distilled water;

[0046] 20mg / mL proteinase K, added separately when using;

[0047] 20% SDS, added separately when using;

[0048] 5M potassium acetate solution, added separately when used.

[0049] The percentages in the above solutions refer to the grams of solute contained in 100 mL of the solution.

[0050] The aforementioned Tris-HCl is a Tris-HCl solution at pH 8.0.

[0051] The above EDTA is an EDTA solu...

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Abstract

The invention relates to a non-toxic extract solution combination GPR.1 for efficiently extracting plant genome DNA and an extraction method based on the extract solution. Specifically include: using SDS extraction buffer containing PVPP and proteinase K and other non-toxic compounds to lyse plant cells to separate chromosomes, denature proteins, and release nucleic acids; then, after adding potassium acetate to remove most of the proteins and polysaccharides, use PEG8000 solution and ethanol / high salt solution to precipitate DNA twice consecutively to remove impurities such as polysaccharides, polyphenols, RNA, NTP, etc., and finally obtain a large amount of high-purity DNA. The method can extract a large amount of high-purity genome DNA from leaves (from young leaves to mature leaves) of various plants, is non-toxic to operators, has little environmental pollution, low cost and short time.

Description

technical field [0001] The invention relates to a method for extracting plant genome DNA and a non-toxic and high-efficiency extracting solution, belonging to the field of biotechnology. Background technique [0002] Isolating DNA that meets the research goals and requirements is the first step in carrying out modern molecular biology research. Compared with animals, in addition to the cell wall, plant cells not only contain polysaccharides, polyphenols and other secondary metabolites that are not easy to separate from DNA, but also the maturity of tissues (such as leaves) used for DNA extraction is often difficult to control. Therefore, the success rate of plant DNA extraction is low and difficult (translated by Chen Zhangliang et al., Angeles et al. Extraction of genomic DNA from the lipid-,polysaccharide-,and polyphenol-rich coconut(Cocos nucifera L.)[J]. Plant MolBiol Rep,2005,23:297a-297i; Ogunkanmi et al.An improved method of extractinggenomic DNA from preserved tissu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003C12Q2521/537
Inventor 高建明桂枝
Owner TIANJIN AGRICULTURE COLLEGE
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