Virus nucleic acid extraction lysis buffer, kit and method

A virus nucleic acid and extraction method technology, applied in the field of tissue sample virus nucleic acid extraction lysate, can solve the problem of polyA solution contaminating aerosol and other problems, and achieve the effects of reducing labor cost and time cost, effective extraction, and less pollution

Pending Publication Date: 2021-05-14
武汉纳磁生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Aiming at the above defects or improvement needs of the prior art, the present invention provides a magnetic bead-based virus nucleic acid extraction method and kit, thereby solving the

Method used

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  • Virus nucleic acid extraction lysis buffer, kit and method
  • Virus nucleic acid extraction lysis buffer, kit and method
  • Virus nucleic acid extraction lysis buffer, kit and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Taking porcine circotype 2 virus vaccine liquid and chicken Newcastle disease virus vaccine liquid as examples, use the magnetic bead method virus nucleic acid extraction kit in Example 1 to compare the extraction with the kits of domestic company A and company B to verify the amplification effect of the extracted products . Among them, sample 1 is a chicken Newcastle disease virus vaccine liquid sample, and sample 2 is a porcine circovirus type 2 virus vaccine liquid sample.

[0049] A kit suitable for a magnetic bead-based viral nucleic acid extraction method, comprising a lysate, a magnetic bead-wash mixture, and an eluent.

[0050] The lysate has a pH value of 8 and contains 3.5mol / L guanidine isothiocyanate, 20mmol / L EDTA, 500mmol / LNaCl, 4wt% Triton, 5wt% polyethylene glycol 8000, and 90mmol / L Tris.

[0051] The pH value of the magnetic bead-washing solution mixture is 7, which contains 75% alcohol, 5 mg / mL magnetic beads, 300 mmol / L NaCl, and the particle diamete...

Embodiment 2

[0062] Taking the mixed oral swab sample of the new crown fake virus liquid as an example, the virus nucleic acid extraction kit of this example was used to compare the extraction effect with the kit of a domestic company C.

[0063] A kit suitable for a magnetic bead-based viral nucleic acid extraction method, comprising a lysate, a magnetic bead-wash mixture, and an eluent.

[0064] The lysate has a pH value of 7.6 and contains 4mol / L guanidine isothiocyanate, 10mmol / L EDTA, 100mmol / LNaCl, 3wt% Triton, 5wt% polyethylene glycol 8000, and 80mmol / L Tris.

[0065] The pH value of the magnetic bead-washing liquid mixture is 7, which contains 75% alcohol, 5 mg / mL magnetic beads, 100 mmol / L NaCl, and the particle diameter of the magnetic beads is 100 nm.

[0066] The eluent contains 10mmol / L Tris-HCl, 1mmol / L EDTA, and the pH is adjusted to 7.

[0067] Apply the kit to carry out automatic extraction of viral nucleic acid, the specific method is as follows:

[0068] 1. Mix the mag...

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Abstract

The invention discloses a magnetic bead-based viral nucleic acid extraction method and a kit. The extraction method comprises the following steps: uniformly mixing a biological sample and a lysis solution to obtain a lysis reaction system; the cleavage reaction system carrying out a reaction, and after the reaction is finished, absorbing nucleic acid through magnetic beads to obtain the magnetic beads adsorbed with the nucleic acid; washing the obtained magnetic beads adsorbed with the nucleic acid, and then eluting the nucleic acid to obtain biological sample nucleic acid; wherein the lysis solution contains a chaotropic agent, a metal chelating agent, sodium ions, a nonionic surfactant and polyethylene glycol. According to the magnetic bead-based virus nucleic acid extraction method, protease K and a PolyA solution are not needed in the nucleic acid release process, and pollution caused by operation is reduced to the maximum extent. The extraction method is convenient and simple to operate, can greatly reduce labor cost and time cost, and is suitable for high-throughput extraction.

Description

technical field [0001] The invention belongs to the field of viral nucleic acid extraction, and more specifically relates to a lysate, a kit and a method for extracting viral nucleic acid from tissue samples. Background technique [0002] With the development of molecular biology, the detection and typing of disease-associated viruses is an important application field of molecular detection, and the efficient and simple extraction and purification of target nucleic acid from biological materials is the key technology of molecular detection. [0003] At present, methods for extracting viral nucleic acid include phenol-chloroform method, boiling method, spin column extraction kit, and magnetic bead method extraction kit. [0004] Among them, although the phenol-chloroform method is more thorough for protein extraction, the operation process is complicated, and the DNA chain is mechanically damaged during repeated extraction and vibration, causing it to degrade. The phenol used...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806C12R1/93
CPCC12Q1/6806C12Q2527/125C12Q2523/308
Inventor 潘超杨小岚宋建军
Owner 武汉纳磁生物科技有限公司
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