Non-toxic extracting solution composition GNR.1 and extracting method for efficiently extracting plant genome DNA

A plant genome, GNR.1 technology, applied in the biological field, can solve problems such as unreported, and achieve the effect of less pollution, no damage to health, and high purity

Active Publication Date: 2018-10-19
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 extracting solution composition GNR.1 and extracting method for efficiently extracting plant genome DNA
  • Non-toxic extracting solution composition GNR.1 and extracting method for efficiently extracting plant genome DNA
  • Non-toxic extracting solution composition GNR.1 and extracting method for efficiently extracting plant genome DNA

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Embodiment 1

[0043] Example 1 Extraction of plant DNA

[0044] 1 Preparation of reagents

[0045] The preparation of the conventional SDS extraction buffer will not be repeated here. The following are the components and concentrations of the GNR.1A, GNR.1D and GNR.1E solutions.

[0046] Solution GNR.1A: 2.7% PEG8000, 0.54M NaCl, 0.11M Tris-HCl (pH 8.0), 0.054M EDTA (pH 8.0), 0.25% Vc (added just before use)

[0047] Solution GNR.1D: 19% PEG8000, 3.7M NaCl

[0048] Solution GNR.1E: 20mM Tris-HCl (pH8.0), 2mM EDTA (pH8.0), 2.5M NaCl

[0049] The 20% SDS described below is a SDS solution with a mass concentration of 20%.

[0050] The percentages in the above solutions refer to the grams of solute contained in 100 mL of the solution.

[0051] 2 Extraction of genomic DNA from 10 representative plants

[0052] (1) Take 150±50 mg of 11 kinds of leaf samples from 10 kinds of plants (see Table 1), grind them into fine powder in liquid nitrogen, quickly transfer them into a 2 mL centrifuge tube...

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Abstract

The invention discloses a non-toxic extracting solution composition GNR.1 and an extracting method for efficiently extracting a plant genome DNA and belongs to the technical field of biology. The extracting method concretely comprises the following steps of pyrolyzing plant cells by utilizing an SDS extracting buffer liquid containing PEG8000 to enable chromosomes to segregate, proteins to be denatured and nucleic acid to be released, after adding potassium acetate to remove a great amount of proteins and polysaccharides, adopting a PEG8000 / high-salt solution and an ethyl alcohol / high-salt solution to perform continuous DNA precipitation for two times to remove impurities including polysaccharides and polyphenols, and obtaining a great quantity of high-purity DNA. By adopting the method, agreat quantity of high-purity genome DNA can be extracted from young leaves or mature leaves of various plants, and the method is non-toxic to an operator, is small in environmental pollution, low incost and short in time and can be widely applied to DNA extraction of the plants.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a method for extracting plant DNA, in particular to a non-toxic extracting solution combination GNR.1 for efficiently extracting plant genomic DNA and an extraction method. Background technique [0002] Isolation of DNA that meets 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. [0003] According to the different surfactants used in the extraction buffer and the final purification and recovery methods of DNA, there are usually four methods for separating DNA from plant tissues (see Table 1), na...

Claims

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Application Information

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