Kit for extracting DNA/RNA of virus through magnetic bead method and using method

A magnetic bead method and kit technology, applied in the field of molecular biology, can solve the problems of cumbersome steps, low purity of nucleic acid DNA/RNA concentration, difficult preservation of nucleic acid, etc.

Inactive Publication Date: 2015-06-03
宝瑞源生物技术(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although nucleic acid separation technology is developing rapidly at present, there are some shortcomings: for example, although the phenol-chloroform extraction method has high nucleic acid purity, it is harmful to the human body and is likely to cause environmental pollution. At the same time, the extraction process requires repeated extractions and multiple tube changes. , the steps are cumbersome, which will cause the loss and contamination of samples
Alkaline lysis method: the nucleic acid extracted by this method is not easy to preserve and cannot be used to extract RNA
The concentration and purity of nucleic acid DNA / RNA extracted by silica gel membrane adsorption column method is low, it is difficult to realize automation, and the throughput per unit time is low

Method used

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  • Kit for extracting DNA/RNA of virus through magnetic bead method and using method
  • Kit for extracting DNA/RNA of virus through magnetic bead method and using method

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Experimental program
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Effect test

preparation example Construction

[0026] Preparation and preservation of serum or plasma liquid samples:

[0027] 1) Serum preparation: Draw 2ml of the subject’s venous blood with a disposable sterile syringe, inject it into a sterile dry glass tube, and let it stand at room temperature for 30-60 minutes, but not more than 4 hours. Centrifuge for 5-10 minutes, aspirate the upper serum and transfer to a sterile centrifuge tube;

[0028] 2) Plasma preparation: Draw 2ml of the subject's venous blood with a disposable sterile syringe, inject it into a glass tube containing disodium ethylenediaminetetraacetic acid (EDTA) or sodium citrate anticoagulant, and immediately turn the glass tube upside down Mix the venous blood and anticoagulant thoroughly, place at room temperature for 5-10 minutes, centrifuge at 1500rpm for 5-10 minutes, absorb the upper layer of plasma and transfer it to a sterile centrifuge tube.

[0029] The prepared serum or plasma should be stored at room temperature for no more than 2 hours, stor...

Embodiment 1

[0030] Embodiment 1: Take Hepatitis B (HBV) National Standard Linear Sensitivity L0-L7 of National Standard Product of China National Institute of Inspection and Quarantine as the sample to be tested, L7 is L6 10-fold dilution gain, concentration is 3-30 IU / ml, use kit of the present invention and The above samples were extracted and detected by fluorescent quantitative PCR.

[0031] 1) Add 300ul of extraction solution I, 60ul of proteinase K, 4ul of nucleic acid sedimentation aid, 20ul of magnetic beads, 200ul of each sensitivity reference product of the national reference plate, and 100ul of extraction solution II into a 1.5ml nuclease-free centrifuge tube, and place at room temperature for 15min without stopping Mix well by inverting to make the magnetic beads and nucleic acid fully contact, centrifuge at 1500rpm instantaneously, place the centrifuge tube on the magnetic stand for 2min, so that the magnetic beads in the tube are adsorbed, and suck the liquid in the tube with...

Embodiment 2

[0032] Example 2: Take hepatitis C (HCV) strongly positive serum sample as the sample to be tested, and dilute it in a 10-fold gradient, use this kit and the inventive method to extract the above sample and perform fluorescence quantitative PCR detection.

[0033] 1) Add 300ul of extraction solution I, 60ul of proteinase K, 4ul of nucleic acid precipitating agent, 20ul of magnetic beads, 200ul of serum samples containing hepatitis C virus, and 100ul of extract solution II into a 1.5ml nuclease-free centrifuge tube. Mix by inverting to make the magnetic beads and nucleic acid fully contact, centrifuge at 1500rpm instantaneously, place the centrifuge tube on the magnetic stand for 2 minutes, so that the magnetic beads in the tube are absorbed, and suck the liquid in the tube with a pipette. 2) Add 550ul of extraction solution III, shake for 5s to suspend the magnetic beads, centrifuge instantaneously, place the centrifuge tube on the magnetic stand for 2min to absorb the magnetic...

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Abstract

The invention discloses a kit for extracting DNA/RNA of a virus by utilizing magnetic beads and a using method. The kit comprises the following eight components: an extracting solution I (cracking), an extracting solution II (binding), an extracting solution III (scrubbing), an extracting solution IV (scrubbing), an extracting solution V (eluting), magnetic bead suspension, a protease K working solution and a nucleic acid settling agent. The kit extraction method comprises the following steps: cracking, binding, scrubbing and eluting. The kit and the extraction method can be used for extracting the DNA/RNA of the viruses of different types of samples, the impurities such as proteins and lipids in the samples are effectively removed, the extracted nucleic acid is high in purity and complete in fragments, and the extraction process is safe and non-toxic.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, in particular to a reagent for extracting virus DNA / RNA by a magnetic bead method and a use method thereof. Background technique [0002] Description of background technology Molecular biology technology can be applied to the research of genetic diseases and the detection of pathogens, and has improved the research on the etiology, pathogenesis, diagnosis and treatment of tumors to the gene molecular level, nucleic acid separation and purification technology It is a basic technique of biochemistry and molecular biology. Nucleic acid is the carrier of genetic information and the material basis of gene expression. Whether it is to study the structure or function of nucleic acid, it is first necessary to isolate the nucleic acid. [0003] Although nucleic acid separation technology is developing rapidly at present, there are some shortcomings: for example, although the phenol-chloroform ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 郭冬冬迟磊杨海侠张琳吕翔
Owner 宝瑞源生物技术(北京)有限公司
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