Kit for extracting viral nucleic acid and extraction method

Abstract

The invention provides a kit for extracting viral nucleic acid and an extraction method. The kit for extracting viral nucleic acid comprises a lysis solution, a magnetic bead suspension, a protease Kdissolving solution, a nucleic acid settling agent solution, a washing solution and an eluent, wherein the lysis solution comprises Tris-fHCL, guanidinium isothiocyanate, sodium N-lauroyl sarcosine, EDTA, TritonX-100, CTAB and Antifoam 204; the washing solution comprises Tris-HCL, PEG6000 and sodium chloride; and the eluent is pure water. According to the kit and the extraction method, the extraction effect of nucleic acid is improved, extraction of viral nucleic acid at room temperature can be realized, gene pollution caused by heating treatment is avoided, in addition, the influence of ethanol on the extraction process is avoided, and compared with a conventional magnetic bead method, the extraction time can be shortened to about 15 minutes, and the extraction process is faster, simplerand more convenient.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a viral nucleic acid extraction kit and extraction method. Background technique [0002] Nucleic acid extraction technology is the basis of virus molecular biology research, and has been widely used in many fields such as pathogenic microorganism detection, clinical disease diagnosis, environmental microorganism detection, food safety detection and forensic identification. During the extraction process, the following principles should be followed: extract as fully as possible, ensure the integrity of the primary structure, avoid contamination of biological macromolecules such as proteins, lipids, and polysaccharides, and shorten and simplify the extraction process as much as possible. A nucleic acid extraction reagent with high extraction efficiency, short time and simple operation has become the key to rapid molecular diagnostic technology. [0003] At present, magnet...

AI Technical Summary

Problems solved by technology

It specifically includes four steps of lysis, adsorption, washing, and elution. Among them, the lysis is mainly to extract the DNA / RNA of the sample to be tested through guanidine isothiocyanate, and the adsorption is usually performed on a magnetic stand. The magnetic beads adsorb DNA / RNA, and the washing is mainly carried out three times. The first two mainly use the buffer whose main components are guanidine hydrochloride and ethanol to remove protein impurities, and the third time mainly use the buffer whose main component is ethanol to remove salt ions. Finally, it is necessary to dry the magnetic beads in a ventilated place, and then use Tris-HCL or aqueous solution to elute the magnetic beads. There are many defects in the existing magnetic bead method. Incubate for 15-20 minutes, with an interval of about 5 minutes during the period. It needs to be inverted 3-4 times, which is very troublesome to operate; the drying process is mainly used to remove residual ethanol, which is not only time-consuming, but also improper operation may easily lead to residual ethanol, affecting PCR amplification. increase rate; in order to accelerate the release and elution of DNA / RNA, heat treatment is usually carried out during the lysis and elution process, which is likely to cause genetic contamination during heating, resulting in false positive test results; the entire extraction time of the existing magnetic bead method needs About 40min, time-consuming is too long, therefore, those skilled in the art are devoted to developing a kind of virus nucleic acid extraction kit and extraction method, make up for existing defect

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