Alcohol-free virus nucleic acid extraction kit based on magnetic bead method

A virus nucleic acid and kit technology, applied in the field of bioengineering, can solve the problems of complex nucleic acid extraction methods, difficult purification, and high extraction requirements, and achieve the effects of significant technological progress, broad application prospects, and simple experimental steps.

Pending Publication Date: 2019-05-07
SHANGHAI KEHUA BIO ENG
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Problems solved by technology

[0004] Aiming at the above-mentioned technical problems in the prior art, the invention provides a kind of alcohol-free virus nucleic acid extraction kit based on the magnetic bead method, and the described alcohol-free virus nucleic acid extraction kit based on the magnetic bead method should solve the existing The method of extracting nucleic acid in the technology is complex, the extraction requirements are high, and the technical problems of purification are difficult

Method used

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  • Alcohol-free virus nucleic acid extraction kit based on magnetic bead method
  • Alcohol-free virus nucleic acid extraction kit based on magnetic bead method
  • Alcohol-free virus nucleic acid extraction kit based on magnetic bead method

Examples

Experimental program
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Effect test

Embodiment 1

[0027] The detection of HBV and HCV virus in the blood plasma of embodiment 1 (comparative research is carried out with the magnetic bead method kit of certain domestic producer)

[0028] (1) Extraction of viral nucleic acid

[0029] 1) Add 320ul of plasma to a 2ml centrifuge tube (add WHO HBV standard with a final concentration of 10IU and WHO HCV standard with a final concentration of 30IU to the negative plasma), add 600μL of lysate, 15μL of solution containing magnetic beads, and 20μL of protease K solution and 5 μL polyA solution, vortex 1min until the sample is fully mixed, then shake and mix for 15min.

[0030] 2) After instantaneous centrifugation, place the centrifuge tube on the magnetic stand and let it stand for 1-2 minutes. After the magnetic beads are completely absorbed, discard the liquid.

[0031] 3) Add 500 μL of buffer solution to remove protein, mix well, and vortex for 1 min.

[0032] 4) After instantaneous centrifugation, place the centrifuge tube on th...

Embodiment 2

[0053] Example 2 Detection of HBV and HCV viruses in serum (compared with Shanghai Kehua Nucleic Acid Extraction Kit (lot: 18076550), refer to the instructions for the operation steps)

[0054] (1) Extraction of viral nucleic acid

[0055] 1) Add 320ul of plasma to a 2ml centrifuge tube (add WHO HBV standard with a final concentration of 10IU and WHO HCV standard with a final concentration of 30IU to the negative plasma), add 600μL of lysate, 15μL of solution containing magnetic beads, and 20μL of protease K solution and 5 μL polyA solution, vortex 1min until the sample is fully mixed, then shake and mix for 15min.

[0056] 2) After instantaneous centrifugation, place the centrifuge tube on the magnetic stand and let it stand for 1-2 minutes. After the magnetic beads are completely absorbed, discard the liquid.

[0057] 3) Add 500 μL of buffer solution to remove protein, mix well, and vortex for 1 min.

[0058] 4) After instantaneous centrifugation, place the centrifuge tube...

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Abstract

The present invention provides an alcohol-free virus nucleic acid extraction kit based on a magnetic bead method. The alcohol-free virus nucleic acid extraction kit comprises a lysate, a magnetic beadsolution, proteinase K, a polyA solution, a deproteinized solution, a washing solution and a buffer solution. The present invention also provides a method for extracting nucleic acid by using the above kit. Firstly, a sample, the lysate and magnetic beads are mixed evenly, then nucleic acid molecules are captured while pyrolysis is conducted, and finally the special alcohol-free deproteinized solution and the washing solution are respectively used to remove impurities of proteins, salt ions and other impurities on surfaces of the magnetic beads. Compared with a conventional virus nucleic acidextraction method, the method is simple in operation steps, does not need single addition of binding liquid, realizes the simultaneous pyrolysis and binding, and co-extracts DNA and RNA; the whole operation process is free of alcohol or isopropyl alcohol, inhibitory substance residues are less, the experimental procedure is simple, and the whole process only takes about 40 minutes; and the method is safe and pollution-free, and free of toxic reagents of chloroform, phenol, etc., and can be operated manually or on an automation platform.

Description

technical field [0001] The invention belongs to the field of bioengineering and relates to a detection kit, in particular to an alcohol-free viral nucleic acid extraction kit based on a magnetic bead method. Background technique [0002] The structure of the virus is relatively simple, consisting of the outer capsid protein and the inner nucleic acid substance. According to the nucleic acid substance, the virus can be divided into DNA virus or RNA virus. 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 acids from biological materials is the key technology of molecular detection. Currently, viral nucleic acid purification technologies are mainly divided into three types, namely traditional method, column membrane method and magnetic bead method. [0003] Traditional nucleic acid purification main...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12N1/06
CPCY02A50/30
Inventor 王金辉方琴张健蒋淼
Owner SHANGHAI KEHUA BIO ENG
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