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A method for selectively extracting dna

A selective, DNA molecule technology, applied in the biological field, can solve the problems of low recovery efficiency, easy pollution, time-consuming and labor-intensive, etc., and achieve the effect of simple raw materials, cheap raw materials, and fast and convenient operation.

Active Publication Date: 2020-05-05
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the selective extraction of DNA fragments of different lengths is usually carried out by gel electrophoresis recovery method, which is not only time-consuming and laborious, but also has many disadvantages such as easy pollution and low recovery efficiency.

Method used

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  • A method for selectively extracting dna
  • A method for selectively extracting dna

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Disperse 0.5mg surface lysine-modified diatomaceous earth particles in 100μl aqueous solution containing 2000bp plasmid DNA and 100bp linear double-stranded DNA molecules (concentrations are both 25ng / μl), and use NaCl to adjust the system Na + The concentration is 0.4mol / L. Adjust the pH of the solution to 5.0, and shake the shaker for 20 minutes after mixing. Centrifugation at 6000 rpm was used to separate diatomaceous earth particles with DNA adsorbed. In addition, 0.75 mg of diatomaceous earth particles modified with surface lysine were dispersed in the adsorption supernatant, and the pH of the solution was adjusted to 4.0. After mixing, the shaker was shaken for 20 minutes, and the diatomaceous earth particles that had adsorbed DNA for the second time were separated by centrifugation at 6000 rpm.

[0025] The DNA-adsorbed diatomaceous earth particles separated by two centrifugation were separately dispersed in a pH=8.0 aqueous solution, and the shaker was shaken for ...

Embodiment 2

[0027] Disperse 1.0 mg of diatomaceous earth particles modified with surface lysine in 100 μl of an aqueous solution containing 2000 bp plasmid DNA and 100 bp linear double-stranded DNA molecules (with a concentration of 25 ng / μl), and use KCl to adjust the K in the system. + The concentration is 0.5mol / L. Adjust the pH of the solution to 4.0, and shake the table for 30 minutes after mixing. Centrifugation at 6000 rpm was used to separate diatomaceous earth particles with DNA adsorbed. In addition, 1.0 mg of diatomaceous earth particles modified with surface lysine were dispersed in the adsorption supernatant, and the pH of the solution was adjusted to 3.0. After mixing, the shaker was shaken for 20 minutes, and the diatomaceous earth particles that had adsorbed DNA for the second time were separated by centrifugation at 6000 rpm.

[0028] Disperse the DNA-adsorbed diatomaceous earth particles separated by centrifugation twice in an aqueous solution of pH=11.0, mix well and shak...

Embodiment 3

[0030] Disperse 0.5 mg of diatomaceous earth particles modified with surface lysine in 100μl of aqueous solution containing 1000bp and 100bp linear double-stranded DNA molecules (at a concentration of 25ng / μl each), adjust the salt concentration in the system to 0.4mol / L, adjust the solution The pH value is 5.0, and the shaker shakes for 20 minutes after mixing. Centrifuge at 6000 rpm to separate diatomaceous earth particles that have adsorbed DNA. In addition, 1.0 mg of diatomaceous earth particles modified with surface lysine were dispersed in the adsorption supernatant, and the pH value of the solution was adjusted to 4.0. After mixing, the shaker was shaken for 20 minutes, and the diatomaceous earth particles that had adsorbed DNA for the second time were separated by centrifugation at 6000 rpm.

[0031] Disperse the DNA-adsorbed diatomaceous earth particles separated by centrifugation twice in a pH=10.0 aqueous solution, mix well, and shake in a shaker for 20 minutes. Cent...

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Abstract

The invention discloses a method for extracting DNA selectively, and belongs to the technical field of biotechnologies. The method comprises the following steps: mixing a solid phase medium after surface modification and a DNA sample solution; adding neutral salt, and enabling the salt concentration in the mixed solution to be 0.4-0.5 mol / L; adjusting the pH value of the mixed solution to be 4-5; conducting oscillation for 20-30 minutes; conducting centrifugal separation; adjusting the pH value of adsorption supernatant fluid to be 3-4; adding the solid phase medium after surface modification; conducting oscillation for 20-30 minutes; conducting centrifugal separation; dispersing the solid phase medium which is separated, and adsorbs DNA molecules of each time in an aqueous solution of which the pH value is 8.0-11.0; conducting oscillation for 20-30 minutes; conducting centrifugal separation to obtain DNA solutions with different length respectively. The method can extract DNA molecules selectively, and has the advantages that the method is high in selecting extraction efficiency, simple and cheap in raw materials, easy to obtain the raw materials, and quick and convenient in operation.

Description

Technical field [0001] The invention belongs to the technical field of biotechnology, and particularly relates to a method for selectively extracting different fragments of DNA by using a solid phase medium based on the salt shielding effect. Background technique [0002] DNA extraction is a basic work in molecular biology, biochemistry, medicine and other fields. With the gradual deepening of the basic and applied research of DNA molecular technology in clinical disease diagnosis, import and export inspection and quarantine, forensic examination, blood quality control and other fields. The selective and purposeful extraction of DNA molecules, especially the selective extraction of small fragments of DNA, has become a difficult problem in the application and promotion of DNA analysis technology. [0003] In 1979, a solid-phase DNA extraction method based on silica was first proposed. At present, the commercially used DNA extraction kits are mainly magnetic bead method, glass bead...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
Inventor 刘玲玲庄家骐杨文胜
Owner JILIN UNIV
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