Nucleic acid extraction kit and nucleic acid extraction method

A nucleic acid extraction reagent and nucleic acid technology, applied in biochemical equipment and methods, microbiological measurement/inspection, DNA preparation, etc., can solve the problems of low purity and concentration of extracted nucleic acid, cumbersome extraction kit steps, poisonous and harmful, etc., to achieve The extraction method is simple, the time cost is shortened, and the effect of saving production and application costs

Pending Publication Date: 2021-08-24
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The present invention provides a nucleic acid extraction kit and a nucleic acid extraction method, which solves the problems that the existing nucleic acid extraction kits have cumbersome steps, take a long time, introduce toxic and harmful organic reagents and other amplification reaction inhibitors, and extract nucleic acid with low purity and concentration question

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  • Nucleic acid extraction kit and nucleic acid extraction method
  • Nucleic acid extraction kit and nucleic acid extraction method
  • Nucleic acid extraction kit and nucleic acid extraction method

Examples

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Embodiment 1

[0072] In this embodiment, the HeLa cell suspension is used as a sample, and the nucleic acid in the sample is extracted. The nucleic acid extraction process of the present embodiment is as follows:

[0073] (1) Add 200 μL of HeLa cell suspension and 200 μL of chelating resin suspension into a 1.5ml centrifuge tube, mix thoroughly by inversion, place at 95°C for 10 minutes, and vortex and shake the mixed solution several times during the process to ensure the chelation Lysis of cells by synthetic resin and specific adsorption of non-nucleic acid impurities;

[0074] (2) Centrifuge the mixed liquid at a speed of 5000rpm for 2 minutes, and draw the supernatant into another new 1.5ml centrifuge tube;

[0075] (3) Add 50 μL of magnetic nanoparticle solution, 20 μL of proteinase K solution, and 250 μL of buffer solution into the centrifuge tube, mix them upside down, and incubate at room temperature for 5 minutes. full adsorption;

[0076] (4) Place the centrifuge tube on the ma...

Embodiment 2

[0086] In this embodiment, the human umbilical vein endothelial cell suspension is used as a sample, and the nucleic acid in the sample is extracted. The nucleic acid extraction process of the present embodiment is as follows:

[0087] (1) Add 200 μL of human umbilical vein endothelial cell suspension and 200 μL of chelating resin suspension into a 1.5ml centrifuge tube, mix thoroughly by inversion, place at 95°C for 10 minutes, and vortex the mixed solution several times during the process The treatment ensures the lysis of cells by the chelating resin and the specific adsorption of non-nucleic acid impurities;

[0088] (2) Centrifuge the mixed liquid at a speed of 5000rpm for 2 minutes, and draw the supernatant into another new 1.5ml centrifuge tube;

[0089] (3) Add 50 μL of magnetic nanoparticle solution, 20 μL of proteinase K solution, and 250 μL of buffer solution into the centrifuge tube, mix them upside down, and incubate at room temperature for 5 minutes. full adsor...

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Abstract

The invention relates to a nucleic acid extraction kit and a nucleic acid extraction method, the nucleic acid extraction kit comprises a magnetic nanoparticle solution, a chelate resin suspension, a protease K solution, a washing solution and an eluent, the magnetic nanoparticles are effectively modified and pretreated by chitosan, and the proper pH value of the solution is controlled to adsorb and desorb nucleic acid; the chelating resin has dual functions of mechanical lysis of cells and specific adsorption of non-nucleic acid impurities, the washing liquid is used for washing magnetic nanoparticles and absorbing and discarding protein, lipid and cell debris in a solution, and the eluent is used for fully releasing nucleic acid adsorbed on the magnetic nanoparticles to obtain a nucleic acid solution. The nucleic acid extraction process is convenient and fast, toxic and harmful reagents such as lysate, high chaotropic salt and organic solvent or amplification reaction inhibitors are not needed, the extraction process is safe, simple and convenient, the purity and concentration of the extracted nucleic acid are improved, and the obtained nucleic acid solution can be directly applied to subsequent PCR.

Description

technical field [0001] The invention relates to the technical field of nucleic acid extraction, in particular to a nucleic acid extraction kit and a nucleic acid extraction method. Background technique [0002] Nucleic acid is the most basic substance in all biomolecules, and high-quality extraction of nucleic acid is also one of the most critical steps in molecular biology. Nucleic acid extraction can be roughly divided into three steps: cell lysis, removal of impurities, and nucleic acid purification. [0003] The premise of nucleic acid extraction is to release it from cells or other biological substances. Common cell lysis methods include mechanical lysis, guanidinium salt lysis, alkali lysis, CTAB lysis, phenol extraction, etc. Among them, guanidinium salt lysis, alkali lysis, CTAB lysis, phenol extraction, etc. will inevitably use toxic organic solvents and the steps are cumbersome, time-consuming, and extraction efficiency is low, while the mechanical lysis of cells ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013C12Q2521/537C12Q2527/125C12Q2523/308
Inventor 弥胜利赵笑宇李想杨伟豪黄嘉骏
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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