Virus nucleic acid extraction kit based on nano magnetic beads

A viral nucleic acid and nano-magnetic bead technology, which is used in the determination/inspection of microorganisms, DNA preparation, recombinant DNA technology, etc., can solve the problems of difficult automation of lysis, high price, and poor RNA extraction effect, so as to improve the detection rate of samples. , Extract the effect of convenient operation and simple operation

Pending Publication Date: 2021-03-09
BEAVERNANO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, many virus magnetic bead method nucleic acid extraction kits are expensive, use toxic and harmful reagents such as β-mercaptoethanol, and the splitting and combining steps are not easy to automate, and the effect on RNA extraction is poor.

Method used

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  • Virus nucleic acid extraction kit based on nano magnetic beads
  • Virus nucleic acid extraction kit based on nano magnetic beads
  • Virus nucleic acid extraction kit based on nano magnetic beads

Examples

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specific example 1

[0032] An embodiment of the DNA virus nucleic acid extraction method in the serum / plasma sample of the present invention comprises the following steps:

[0033] 1. Sample preparation: take the HBV quality control product as the representative sample of DNA virus, dilute it with negative plasma, and dilute the negative serum to 10 5 pc / mL, as the sample to be tested.

[0034] 2. Viral DNA extraction:

[0035] (1) Take a new 1.5mL centrifuge tube, add 200μL of the sample to be tested, then add 40μL of proteinase K, 400μL of lysate, 1μL of transfer RNA, 200μL of isopropanol and 20μL of nano-magnetic beads, adjust the appropriate speed and vortex to mix well , and heat the centrifuge tube at 55°C for 15 minutes (invert several times every 5 minutes). Then place the centrifuge tube on a magnetic separator and let it stand for 1 min, and suck off the supernatant with a pipette.

[0036] (2) Add 600 μL of washing solution I, slowly blow the magnetic beads 10 times with a pipette o...

specific example 2

[0049] An embodiment in which the kit of the present invention can cooperate with an automatic nucleic acid extractor to quickly and high-throughput extract viral RNA in a sample includes the following steps:

[0050] 1. Take the new coronavirus N gene pseudovirus as a representative sample of RNA virus, add virus preservation solution and dilute to 10 5 pc / mL, as the sample to be tested.

[0051] 2. Use Tianlong Technology Co., Ltd. NP968 Nucleic Acid Automatic Extractor, and the instructions for adding reagents to the wells are as follows:

[0052] Wells 1 and 7: 200 μL of sample, 40 μL of proteinase K, 400 μL of lysate, 1 μL of transfer RNA, 200 μL of isopropanol;

[0053] Wells 2 and 8: 600 μL of washing solution I;

[0054] Wells 3 and 9: 600 μL of washing solution II;

[0055] Wells 4 and 10: 600 μL of washing solution II;

[0056] Wells 5 and 11: 20 μL of magnetic beads;

[0057] Wells 6 and 12: 100 μL of Nuclease-free Water.

[0058] 3. The program setting of NP9...

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Abstract

The invention discloses a virus nucleic acid extraction kit based on nano magnetic beads. The lysis liquid comprises guanidine thiocyanate/guanidine hydrochloride, a nonionic surfactant, tris (hydroxymethyl) aminomethane hydrochloride and ethylenediamine tetraacetic acid, and the nonionic surfactant comprises one or more of triton X-100, Tween 20 and ethyl phenyl polyethylene glycol. The binding liquid is isopropanol, the volume ratio of isopropanol to pyrolysis is 1: 1, and the magnetic beads are silicon oxide hydroxyl nano magnetic beads. According to the kit, a nano magnetic bead extractionmethod is adopted, isopropanol is used as a binding liquid, making sample splitting, binding of nucleic acid and magnetic beads to be done in one step. The steps are simple, convenient and easy to implement, and the automatic extraction process of virus nucleic acid can be completed quickly at high flux within 9 min. The kit disclosed by the invention can be used for extracting RNA and DNA of viruses in sample, such as plasma, serum, nasopharynx swabs, urine, secretion liquid, virus concentrate, cell culture liquid supernatant and virus preservation liquid.

Description

technical field [0001] The invention relates to the technical field of nucleic acid extraction and purification, in particular to a virus nucleic acid extraction kit based on nano magnetic beads. Background technique [0002] At present, the methods for extracting nucleic acid mainly include the traditional phenol / chloroform organic liquid phase extraction method, alkali cracking method, cetyltrimethylammonium bromide (CTAB) extraction method, non-traditional chelating resin method, and glass powder adsorption method. , Silica gel membrane adsorption column method. The traditional phenol / chloroform organic liquid phase extraction method has toxic organic solvents that will cause harm to the human body and has many steps and cumbersome operations; the alkaline lysis method is mostly used for DNA extraction, and the product contains impurities, which is not conducive to downstream detection experiments. In addition, RNA The extraction effect is also poor. The silica gel memb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/6806
CPCC12N15/1013C12Q1/6806C12Q2523/10C12Q2523/308C12Q2527/125C12Q2563/143C12Q2563/155
Inventor 任辉付监贵
Owner BEAVERNANO TECH
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