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DNA extraction adsorption liquid, kit and DNA extraction method based on super paramagnetic iron oxide nanoparticles

A nanoparticle and superparamagnetic technology, applied in the field of DNA extraction, can solve problems such as the inability to meet the needs of efficient and accurate scientific research, the limitation of the promotion of basic biological sequencing technology, and the unreasonable DNA extraction kits. The effect of high and low extraction cost

Pending Publication Date: 2020-04-28
AGRI GENOMICS INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the existing extraction kits use different magnetic particles and adsorbents, and the extraction effects are very different, and their prices are also very expensive.
At present, the most widely used imported Beckman AMPure XP kit has a price as high as 216.67 yuan / ml; other domestic magnetic particle extraction kits have low extraction efficiency and cannot meet the needs of efficient and accurate scientific research, and their prices are also at 100 yuan / ml about ml
Expensive DNA extraction kits are very unreasonable, and this situation also limits the development of basic biology and the promotion of sequencing technology in my country to a certain extent

Method used

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  • DNA extraction adsorption liquid, kit and DNA extraction method based on super paramagnetic iron oxide nanoparticles
  • DNA extraction adsorption liquid, kit and DNA extraction method based on super paramagnetic iron oxide nanoparticles
  • DNA extraction adsorption liquid, kit and DNA extraction method based on super paramagnetic iron oxide nanoparticles

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Experimental program
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Effect test

Embodiment 1

[0037] The brief principle and steps of DNA extraction in this embodiment are as follows: figure 1 shown; the specific steps are as follows:

[0038] Preparation of adsorption solution

[0039] The adsorption solution in this example is composed of PEG, NaCl, MgCl 2 Composed of three ingredients. The specific configuration method of the storage solution is as follows:

[0040] Storage solution A: Weigh 14.61g of NaCl, add 40ml of ultrapure water, place it in a rotator at room temperature and turn it over to dissolve. After most of the white particles are dissolved, let it stand at room temperature for 1-6 hours to fully dissipate the bubbles in the tube, and the tube wall liquid The drops are fully polymerized, which is convenient and accurate to make the volume, and finally make the volume to 50ml, and then turn it over again to dissolve. After completely dissolving, use a 0.22 μm microporous membrane filter to complete the filtration, and store at 4°C for later use.

[...

Embodiment 2

[0068] The brief principle and steps of DNA extraction in this embodiment are as follows: figure 1 shown; the specific steps are as follows:

[0069] Preparation of adsorption solution

[0070] The adsorption solution in this example is composed of PEG, NaCl and MgCl2. The specific configuration method is as in Example 1.

[0071] Preparation of working fluid for magnetic particles

[0072] The magnetic particle working solution is composed of superparamagnetic nanoparticles and an adsorption solution. The carboxyl-modified coated magnetic particles used in this example can be purchased directly from a biochemical reagent store, and its storage solution is 20% ethanol solution. The specific steps for configuring the magnetic particle working mixture are as follows:

[0073] Incubate the magnetic particle storage solution in advance. In order to ensure the accuracy of the volume taken, it is necessary to take out the magnetic particle storage solution from 4°C in advance an...

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Abstract

The present invention provides DNA extraction adsorption liquid, a kit and a DNA extraction method based on super paramagnetic iron oxide nanoparticles, and belongs to the technical field of DNA extraction. The adsorption liquid comprises polyethylene glycol, sodium chloride and magnesium chloride; concentration of the polyethylene glycol is 100-250 mg / ml; concentration of the sodium chloride is 1.25-2.75 mol / L; and concentration of the magnesium chloride is 10-20 mmol / L. The extraction method comprises the following steps: 1) mixing the super paramagnetic iron oxide nanoparticles with the adsorption liquid; 2) mixing DNA samples to be extracted with working liquid, performing magnetic separation after adsorption extraction, and collecting the super paramagnetic iron oxide nanoparticles adsorbing with the DNA; and 3) eluting the DNA-adsorbed super paramagnetic iron oxide nanoparticles, performing magnetic separation, and collecting liquid phase components to obtain the extracted DNA. The adsorption liquid has simple components and high adsorption efficiency; and the extraction method has fast extraction speed and low cost.

Description

technical field [0001] The invention belongs to the technical field of DNA extraction, and in particular relates to a superparamagnetic nanoparticle-based DNA extraction adsorption solution, a kit and a DNA extraction method. Background technique [0002] DNA samples and DNA-based experiments are the most common and fundamental in the life sciences. High-concentration, high-purity DNA extraction has become a key step in molecular biology experiments, and it is also the basic first step for many downstream experiments. Traditional methods for extracting DNA mainly rely on phenol-chloroform or column purification. However, the use of toxic chemical reagents and low extraction efficiency make these two methods unable to meet the needs of modern scientific research. [0003] In the past few decades, magnetic particles have been widely used in nucleic acid extraction. The magnetic particle extraction reagent is mainly composed of two parts: magnetic particles and adsorption so...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013Y02P10/20
Inventor 张玉波李秋佳
Owner AGRI GENOMICS INST CHINESE ACADEMY OF AGRI SCI