Non-toxic extract composition GPR.1 capable of efficiently extracting plant genome DNA and extraction method

A plant genome and extraction technology, applied in the biological field, can solve problems that have not been reported, and achieve the effects of low pollution, high purity, and low cost

Active Publication Date: 2018-11-20
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 composition GPR.1 capable of efficiently extracting plant genome DNA and extraction method
  • Non-toxic extract composition GPR.1 capable of efficiently extracting plant genome DNA and extraction method
  • Non-toxic extract composition GPR.1 capable of efficiently extracting plant genome DNA and extraction method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Preparation of extract:

[0042] The preparation of the conventional SDS extraction buffer will not be repeated. The following is the composition of the 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 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 using.

[0049] The percentage in the above solution refers to the grams of solute contained in 100 mL of the solution.

[0050] The above Tris-HCl is a Tris-HCl solution with pH 8.0.

[0051] The above-mentioned EDTA is an EDTA solution of pH 8.0.

[0052] Extract...

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Abstract

The invention relates to a non-toxic extract composition GPR.1 capable of efficiently extracting plant genome DNA and an extraction method based on the extract. The method specifically comprises the following steps: lysing plant cells with an SDS extraction buffer containing PEG8000, PVPP (Polyvinylpolypyrrolidone) and protease K, performing chromosome segregation and protein denaturation, and releasing nucleic acids; adding potassium acetate to remove most of the proteins and polysaccharides, and continuously precipitating the DNA twice by adopting a PEG8000 / high-salt solution and an ethanol / high-salt solution so as to remove the polysaccharides, polyphenol, RNA, NTP and other impurities, thereby finally obtaining lots of high-purity DNA. According to the method disclosed by the invention, lots of high-purity genome DNA can be extracted from leaves (young leaves or mature leaves) of multiple plants, and the method is non-toxic to the operator, small in environmental pollution, low incost and short in time.

Description

Technical field [0001] The invention relates to a method for extracting plant genomic DNA and a non-toxic 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 modern molecular biology research. Compared with animals, in addition to cell walls, plant cells not only contain polysaccharides, polyphenols and other secondary metabolites that are not easily separated 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., 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 tissues of Capsicum annuum for PCR amplifica...

Claims

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Application Information

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