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Nucleic acid extraction kit and nucleic acid extraction method

A nucleic acid extraction reagent and extraction method technology, applied in the field of nucleic acid separation, purification or preparation, can solve the problem of low efficiency of extracting nucleic acid molecules, and achieve the effects of reducing the use of chemical reagents, good dispersibility, and high-efficiency adsorption

Active Publication Date: 2018-12-21
SUZHOU BAIYUAN GENT CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Therefore, the technical problem to be solved in the present invention is to overcome the low efficiency of extracting nucleic acid molecules by using magnetic solid phase extraction technology in the prior art

Method used

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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

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] This embodiment provides a magnetic nanoparticle (PDA@Fe 3 o 4 ) preparation method, comprising the following steps:

[0062] (1) with FeCl 3 ·6H 2 O as raw material, magnetic Fe synthesized by hydrothermal method 3 o 4 Nanoparticles

[0063] FeCl 3 ·6H 2O (1.35 g) was dissolved in ethylene glycol (40 mL) and a clear solution was formed by sonication. Then, sodium acetate (3.6 g) and polyethylene glycol 10000 (1.0 g) were added to the solution, and the mixture was vigorously stirred until a homogeneous dark yellow solution was obtained. The solution was put into a polytetrafluoroethylene high pressure reactor and heated at 200° C. for 48 hours. The product was washed several times with ethanol and water, and dried under a nitrogen atmosphere at 60 °C for 10 h to obtain carboxyl-modified magnetic Fe 3 o 4 Nanoparticles, spare;

[0064] (2) Dopamine hydrochloride (200 mg) was mixed with 0.12 g of Tris-HCl buffer (10 mM, pH 8.5) and 100 ml of water to prepare a...

Embodiment 2

[0079] This embodiment provides a nucleic acid extraction kit, including a nucleic acid binding solution and a magnetic adsorbent:

[0080] The nucleic acid binding solution includes 20% (w / v) polyethylene glycol, 4mol / L sodium chloride, and the pH of the nucleic acid binding solution is 2;

[0081] The magnetic adsorbent is the magnetic nanoparticles prepared in Example 1 (PDA@Fe 3 o 4 ).

Embodiment 3

[0083] This embodiment provides a nucleic acid extraction method, using the nucleic acid extraction kit provided in Example 2, the extraction method includes the following steps:

[0084] (1) Cleavage of DNA in human whole blood samples

[0085] ①Put 300μl EDTA-K2 (EDTA dipotassium) anticoagulated blood into a 1.5ml centrifuge tube with a micropipette. Afterwards, 300 μl of sterile deionized water was added to the tube to disrupt the erythrocyte membrane. After repeated mixing several times, the mixture was centrifuged at 12000rpm / min for 3 minutes, and the supernatant was discarded to obtain a precipitate;

[0086] ② Add buffer CL (300 μl) to the precipitate, and mix the mixture several times. The mixture was centrifuged at 12000 rpm / min for 1 minute, and the supernatant was discarded;

[0087] ③ Prepare proteinase K-buffer FG mixture (1:100) (g / V) and add it to the precipitate. Immediately invert the solution slightly several times until the precipitate is completely dis...

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Abstract

The invention discloses a nucleic acid extraction kit comprising a nucleic acid binding solution and a magnetic adsorbent. The nucleic acid binding solution comprises polyethylene glycol and sodium chloride, and the magnetic adsorbent is magnetic Fe3O4 nano particles with the surfaces covered with polydopamine coatings. The magnetic adsorbent provided by the invention has the superparamagetic nanoparticles, the polydopamine coatings on the surfaces of the nano particles are rich in amino groups and hydroxyl groups, under the salt solution environment provided by the nucleic acid binding solution, nucleic acids can be efficiently adsorbed, the nucleic acid adsorbing efficiency and quality of the existing magnetic nano particles are improved, and the nucleic acids can be extracted efficiently and rapidly; and when the nucleic acid extraction kit is applied to extraction of human whole blood, blood treatment is not needed, and the nucleic acid extraction kit is suitable for gene diagnosis and treatment of clinical diseases. The nucleic acid extraction method provided by the invention uses the nucleic acid extraction kit, the quality and efficiency of extracting of the nucleic acids are improved, and the nucleic acid extraction method is suitable for application and promotion of clinical diagnosis and basic scientific research.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid separation, purification or preparation, and in particular relates to a nucleic acid extraction kit and a nucleic acid extraction method. Background technique [0002] The extraction technology of nucleic acid molecules is a key technology in molecular biology research and a prerequisite technology for a series of molecular biology analyzes such as nucleic acid amplification, DNA fragment ligation, vector construction, and high-throughput sequencing. In addition, nucleic acid amplification technology can be used for plant pathogen detection, food detection and disease diagnosis. Therefore, the extraction of nucleic acid is of great significance. Generally speaking, nucleic acid extraction requires multiple steps. First, biological sample materials such as cells and tissue materials need to be crushed to inactivate nucleases, release nucleic acids, and then remove other tissue or cell compone...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/101Y02A50/30
Inventor 车团结尤崇革徐红李琳李亚鹏
Owner SUZHOU BAIYUAN GENT CO LTD
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