Nucleic acid extraction method based on nano magnetic beads and application of nucleic acid extraction method

A technology of nano-magnetic beads and extraction method, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA preparation, etc., can solve the problems of low extraction purity, complicated operation, poor nucleic acid effect, etc., and achieve convenient extraction operation. , a wide range of effects

Inactive Publication Date: 2019-12-31
中诺(杭州)基因科技有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, magnetic bead nucleic acid extraction methods are mostly used in samples such as blood, tissue, swabs, and urine. The extraction of microorganisms and host DNA in feces has low extraction purity and poor extraction effect, which cannot meet the needs of follow-up experiments at the same time. Some nucleic acid extraction methods also have problems such as complicated operation and poor effect of extracted nucleic acid

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  • Nucleic acid extraction method based on nano magnetic beads and application of nucleic acid extraction method
  • Nucleic acid extraction method based on nano magnetic beads and application of nucleic acid extraction method
  • Nucleic acid extraction method based on nano magnetic beads and application of nucleic acid extraction method

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Embodiment 1

[0024] Example 1 see figure 1 , A nucleic acid extraction method based on nano magnetic beads of the present invention includes the following steps:

[0025] Step 1: Take four 200ul of serum containing hepatitis B virus (HBV virus) and place them in a 2ml centrifuge tube as the experimental group (labeled SP1, SP2, SP3, SP4) as the biological samples to be extracted nucleic acid, and add the lysis Buffer 600ul, by changing the permeability of the cells in the serum, the nucleic acid inside the cell is separated from the nucleoprotein, and the nucleoprotein combines with the reagent in the lysis buffer to form a precipitate, thereby releasing the nucleic acid and forming a lysis solution containing the nucleic acid;

[0026] Step 2: Add 10ul of nano magnetic beads to the lysis solution containing nucleic acid, mix and incubate at room temperature for 5 minutes, place the centrifuge tube on a magnetic stand, magnetically attract for 2 minutes to make the magnetic bead-nucleic acid com...

Embodiment 2

[0039] Example 2 see figure 2 , A nucleic acid extraction method based on nano magnetic beads of the present invention includes the following steps:

[0040] Step 1: Take 1g of feces into the feces collection tube, add 3ml of fecal preservation solution, and mix thoroughly, take 200ul of fecal supernatant, add standard amount (10000 copies) of internal reference strain Sa, add 1ml PBS buffer, vortex After shaking evenly, centrifuge at 500 rpm for 3 minutes, collect the supernatant, centrifuge at 5000g for 3 minutes, discard the supernatant, divide into four aliquots and place them in a 2ml centrifuge tube as the experimental group (and labeled SP1, SP2, SP3, SP4) as Add 600ul of lysis buffer to the biological sample from which nucleic acid is to be extracted. By changing the permeability of the cells in the stool, the nucleic acid inside the cell is separated from the nucleoprotein, and the nucleoprotein combines with the reagent in the lysis buffer to form a precipitate, thereb...

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Abstract

The invention discloses a nucleic acid extraction method based on nano magnetic beads. The nucleic acid extraction method comprises the following steps: step 1, adding a lysis buffer to a biological sample, and then forming a lysis solution containing nucleic acid after splitting and precipitating; step 2, adding nano magnetic beads to form magnetic beads-nucleic acid complex, applying an externalmagnetic field to aggregate the magnetic beads-nucleic acid complex; step 3, removing the lysis solution under the action of the external magnetic field, and adding a washing buffer to wash the magnetic beads-nucleic acid complex; step 4, removing the magnetic field, adding an elution buffer to elute the magnetic beads-nucleic acid complex, and then obtaining the required nucleic acid. The invention also discloses an application of the nucleic acid extraction method. The nucleic acid extraction method is used to extract the nucleic acid from faeces, whole blood, serum and plasma, tissue fluid, swab lotion, tissue homogenate, sputum, urine and saliva. The nucleic acid extraction method of the invention has advantages of realizing automatic, efficient, high-throughput extraction and purification of the nucleic acid, and convenient extraction operations.

Description

【Technical Field】 [0001] The invention relates to the technical field of nucleic acid extraction engineering, in particular to the technical field of a nucleic acid extraction method and application based on nano magnetic beads. 【Background technique】 [0002] Biological samples are important materials for analyzing biological gene characteristics. Common biological samples include whole blood, tissue homogenate, swabs, urine, feces, sputum and other samples. Stool, as a new non-invasive sample, is widely used in molecular genetics, intestinal disease diagnosis and other fields. Stool contains many samples with genetic information, such as gastrointestinal bleeding, intestinal flora DNA, DNA from food residue samples, and DNA from exfoliated cells. [0003] Nanomagnetic beads use superparamagnetic nanospheres as carriers, which can quickly separate and purify DNA or RNA, and can be automated, safe and fast, and can reduce the harm of organic solvents to the human body. It can be u...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013C12Q2527/125
Inventor 李强朱罗罗徐威张辉林清源
Owner 中诺(杭州)基因科技有限责任公司
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