Method for extracting plant tissue culture seedling genome DNA (Deoxyribose Nucleic Acid)

A plant tissue culture and genome technology, applied in the field of molecular biology, can solve the problems of increased genomic DNA contamination, loss of natural characteristics, time-consuming and labor-intensive in the extraction process, and achieves the protection of natural activity, shortening extraction time, and convenient process. Effect

Inactive Publication Date: 2014-09-03
INST OF DAFENG MARINE IND NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] There are many methods for plant genomic DNA extraction, most of which are cumbersome and time-consuming.
The lengthy extraction process not only increases the possibility of contamination of genomic DNA, which affects its purity, but also may reduce activity and lose its natural properties
So far, there is no technical method for quick and easy extraction of plant genomic DNA

Method used

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  • Method for extracting plant tissue culture seedling genome DNA (Deoxyribose Nucleic Acid)
  • Method for extracting plant tissue culture seedling genome DNA (Deoxyribose Nucleic Acid)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Select 0.5 g of fresh Pinellia tissue-cultured leaf tissue and put it into a commercial EP tube (specification 1.5 mL); add 1000 μL of SDS and CTAB mixed extract with a volume ratio of 2:1 after preheating with warm water at 65 °C, Shake for 5 minutes; use the tip of a pipette (200 μL) to insert into the bottom of the above EP tube, slowly rotate clockwise to pick up the filamentous viscous substance, which is the obtained genomic DNA, and dissolve it in 50 μL pure water; take the dissolved 3 μL of the final solution was carried out for 1% agarose electrophoresis detection, and the results were as follows figure 1 shown.

Embodiment 2

[0037] Select 0.5g fresh leaf tissue of Bletilla striata tissue culture seedlings at -20°C, put it into a commercial EP tube (1.5mL size); add 65°C warm water to preheat, and mix SDS and CTAB with a volume ratio of 3:1 for extraction Mix 1000 μL of solution for 5 minutes; use the tip of a pipette (200 μL) to insert into the bottom of the above-mentioned EP tube, slowly rotate clockwise to pick up the filamentous viscous substance, which is the obtained genomic DNA, and dissolve it in 50 μL of pure water ; Take 5 μL of the dissolved solution, and carry out 1% agarose electrophoresis detection, the result is as follows: figure 2 shown.

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Abstract

The invention discloses a method for extracting plant tissue culture seedling genome DNA (Deoxyribose Nucleic Acid). The method comprises the following steps: treating a certain amount of leaf tissue of a plant tissue culture seedling, putting into a centrifuge tube, subsequently adding a mixing extraction liquid of SDS (Sodium Dodecyl Sulfate) and CTAB (Cetyltrimethyl Ammonium Bromide) which are preheated by warm water into the centrifuge tube, shaking to mix, further stretching a head of a pipette to the bottom of the centrifuge tube, slowly rotating in a same direction to pick a filiform sticky substance, that is, the obtained genome DNA, finally dissolving the genome DNA into a proper amount of water, and taking a small amount of the water for detection. The method disclosed by the invention is simple to operate, the time for extracting the genome DNA is shortened, the experiment process is simplified, and the natural activity of the genome DNA is effectively protected.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a method for rapidly extracting and detecting herbal plant genome DNA. Background technique [0002] As the carrier of genetic information and the most basic genetic material, genomic DNA plays an important role in genetic variation and metabolic regulation, and is the main research object of molecular biology and genetic engineering. The extraction of genomic DNA is usually used to construct genomic library, Southern hybridization (including RFLP) and PCR to isolate genes, etc. Whether it is to study the structure and function of plant DNA, or to carry out research on the transformation and transduction of exogenous DNA, the first thing to do is to extract natural high-molecular-weight DNA from plant tissues. The genomic DNA extraction methods of different plants are different; different types or different tissues of the same type have different separation methods bec...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 张文涛曹言海陈集双覃佳佳张本厚金磊磊
Owner INST OF DAFENG MARINE IND NANJING UNIV OF TECH
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