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Kit for viral nucleic acid extraction and extraction method thereof

A virus nucleic acid and kit technology, applied in the direction of DNA preparation, recombinant DNA technology, etc., can solve the problems of cumbersome steps, long time-consuming, low extraction rate, etc., and achieve the effect of short time, shortened time, and rapid extraction

Inactive Publication Date: 2019-11-19
广州欣源生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is usually a challenge to isolate high-purity and complete viral nucleic acid. In addition, it is difficult to obtain clinical virus samples, and a large number of samples will increase the suffering of patients.
The traditional virus nucleic acid purification method first uses denaturant, proteinase K and other cleavage solutions to release the nucleic acid, then extracts it with phenol and chloroform, precipitates it with isopropanol, and removes the supernatant to obtain the nucleic acid; the method is cumbersome, time-consuming, and the extraction rate low and poor repeatability
In addition, commercial viral nucleic acid extraction kits are relatively expensive

Method used

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  • Kit for viral nucleic acid extraction and extraction method thereof
  • Kit for viral nucleic acid extraction and extraction method thereof
  • Kit for viral nucleic acid extraction and extraction method thereof

Examples

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Embodiment 1

[0036] This embodiment provides a kit for virus nucleic acid extraction, which includes a lysis solution, a protein-removing solution, an adsorption promoting solution, a washing solution, an eluent, and a purification column; the lysis solution is made of 3mol / L guanidine isothiocyanate , The volume concentration is 0.6% TritonX-100, 30mmol / L Tris-HCl and 30mmol / L urea, the pH of the lysate is 4.3; the protein-removing solution is composed of 5mol / L guanidine hydrochloride and 55% ethanol by volume. The pH value of the protein-removing solution is 6.5; the adsorption promoting solution contains isopropanol and acetic acid aqueous solution, and the volume concentration of acetic acid in the acetic acid aqueous solution is 1%; the washing solution is an aqueous solution containing ethanol and sodium chloride, and the concentration of ethanol in the washing solution is greater than 95%, the concentration of sodium chloride in the washing solution is 20mmol / L, the pH of the washing...

Embodiment 2

[0044] This embodiment provides a kit for virus nucleic acid extraction, which includes a lysis solution, deproteinization solution, adsorption promotion solution, washing solution, eluent and purification column; the lysis solution is made of 5mol / L guanidine isothiocyanate , The volume concentration is 0.8% TritonX-100, 40mmol / L Tris-HCl and 40mmol / L urea, the pH of the lysate is 4.4; the deproteinized solution is composed of 5.5mol / L guanidine hydrochloride and 53% ethanol , The pH value of the protein-removing solution is 6.4; the adsorption promoting solution contains isopropanol and acetic acid aqueous solution, the volume concentration of acetic acid in the acetic acid aqueous solution is 1%; the washing solution is an aqueous solution containing ethanol and sodium chloride, and the volume of ethanol in the washing solution The concentration is greater than 95%, the concentration of sodium chloride in the washing solution is 20 mmol / L, and the pH value of the washing solu...

Embodiment 3

[0047] This embodiment provides a kit for virus nucleic acid extraction, which includes a lysis solution, deproteinization solution, adsorption promotion solution, washing solution, eluent and purification column; the lysis solution is made of 7mol / L guanidine isothiocyanate , The volume concentration is composed of 1% TritonX-100, 50mmol / L Tris-HCl and 50mmol / L urea, the pH value of the lysate is 4.6; the protein-removing solution is composed of 6mol / L guanidine hydrochloride and 59% ethanol. The pH value of the protein-removing solution is 6.6; the adsorption promoting solution contains isopropanol and acetic acid aqueous solution, and the volume concentration of acetic acid in the acetic acid aqueous solution is 1%; the washing solution is an aqueous solution containing ethanol and sodium chloride, and the volume concentration of ethanol in the washing solution >95%, the concentration of sodium chloride in the washing solution is 20mmol / L, and the pH value of the washing solu...

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Abstract

The invention discloses a kit for viral nucleic acid extraction and an extraction method thereof, and belongs to the technical field of nucleic acid extraction. The kit for the viral nucleic acid extraction comprises a lysis solution, a protein removal solution, an absorption promoting solution, a scrubbing solution, an eluant and a purification column, wherein the lysis solution is prepared from3-7mol / L guanidinium isothiocyanate, TritonX-100 with the volume concentration of 0.6%-1%, 30-50mmol / L Tris-HCl and 30-50mmol / L urea, and a pH value of the lysis solution is 4.3-4.6; and in addition,components and content of the protein removal solution, the absorption promoting solution and the scrubbing solution are limited, the kit is cheap, and the extraction method of the kit for the viral nucleic acid extraction can efficiently and stably extract viral nucleic acid.

Description

Technical field [0001] The invention belongs to the technical field of nucleic acid extraction, and particularly relates to a kit for nucleic acid extraction of a virus sample with a small volume of less than 500 μL and an extraction method thereof. Background technique [0002] Viruses not only have potential pathogenicity, but also play an important role in molecular biology and biomedical research. Isolation of high-purity and complete viral nucleic acid is usually a challenge. In addition, it is difficult to obtain clinical virus samples, and a large number of samples will increase the suffering of patients. The traditional virus nucleic acid purification method first uses denaturant, proteinase K and other cleavage solutions to release the nucleic acid, and then extracts with phenol and chloroform, precipitates with isopropanol, and removes the supernatant to obtain the nucleic acid; the method is complicated, time-consuming, and extraction rate. Low and poor repeatability....

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/101
Inventor 刘超武何修业
Owner 广州欣源生物科技有限公司
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