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Magnetic bead method virus nucleic acid extraction kit and use method thereof

A technology of virus nucleic acid and magnetic bead method, which is applied in the medical field, can solve the problems of high cost and low efficiency, and achieve the effect of reducing harm and saving cost

Pending Publication Date: 2022-05-06
江苏迅睿生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the chromatography column nucleic acid extraction technology requires the use of centrifugal columns and automatic centrifugal equipment, the cost is high. At present, the chromatography column nucleic acid extraction is still mainly manual extraction, and the efficiency is low.

Method used

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  • Magnetic bead method virus nucleic acid extraction kit and use method thereof
  • Magnetic bead method virus nucleic acid extraction kit and use method thereof
  • Magnetic bead method virus nucleic acid extraction kit and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Influenza B virus nucleic acid extraction

[0039] The portable steps of the present invention are as follows:

[0040] Dilute influenza B virus 1000 times and 10000 times with negative throat swab samples, take 200 μL respectively and put them in centrifuge tubes, add 700 μL lysis solution, 20 μL 20-50 mg / ml magnetic bead solution, lyse and mix at room temperature for 5 minutes, and put the centrifuge tube Place on a magnetic stand, absorb magnetically for 30s, and discard the supernatant;

[0041] Wherein, the components in the lysate are respectively: 0.25M Tris-HCl, 3.17M guanidine thiocyanate, 25mmEDTA, 3.6% PEG-6000, 2%SDS, 6%Triton-100, 9.5% isopropanol, The rest is water

[0042] Add 1ml of washing solution Ⅰ to the centrifuge tube, mix for 1min, place the centrifuge tube on the magnetic stand, absorb magnetism for 30s, and discard the supernatant;

[0043] Wherein, the components in the washing liquid I are: 3M guanidine hydrochloride, 0.25M Tris-H...

Embodiment 2

[0057] Example 2 Extraction of adenovirus nucleic acid

[0058] The portable steps of the present invention are as follows:

[0059]1) Dilute adenovirus 1000 times and 10000 times with negative stool samples, take 200 μL respectively and put them in centrifuge tubes, add 700 μL lysate, 20 μL 20-50 mg / ml magnetic bead solution, lyse and mix at room temperature for 5 minutes, place the centrifuge tube Place on a magnetic stand, absorb magnetism for 30s, and discard the supernatant;

[0060] Wherein, the components in the lysate are respectively: 0.25M Tris-HCl, 3.17M guanidine thiocyanate, 25mmEDTA, 3.6% PEG-6000, 2%SDS, 6%Triton-100, 9.5% isopropanol, The rest is water

[0061] 2) Add 1ml of washing solution Ⅰ to the centrifuge tube, mix for 1min, place the centrifuge tube on the magnetic stand, absorb the magnet for 30s, and discard the supernatant;

[0062] Wherein, the components in the washing liquid I are: 3M guanidine hydrochloride, 0.25M Tris-HCl, pH adjusted to 6.5 ...

Embodiment 3

[0076] Example 3 Influenza B virus nucleic acid extraction

[0077] The portable steps of the present invention are as follows:

[0078] 1) Dilute influenza B virus 1000 times with negative saliva samples, take 200 μL and place it in a centrifuge tube, add 700 μL of lysate, 20 μL of 20-50 mg / ml magnetic bead solution, lyse and mix for 5 minutes at room temperature, and place the centrifuge tube on a magnetic stand on, absorb magnetically for 30s, and discard the supernatant;

[0079] Wherein, the components in the lysate are respectively: 0.25M Tris-HCl, 3.17M guanidine thiocyanate, 25mmEDTA, 3.6% PEG-6000, 2%SDS, 6%Triton-100, 9.5% isopropanol, The rest is water

[0080] 2) Add 1ml of washing solution Ⅰ to the centrifuge tube, mix for 1min, place the centrifuge tube on the magnetic stand, absorb the magnet for 30s, and discard the supernatant;

[0081] Wherein, the components in the washing liquid I are: 3M guanidine hydrochloride, 0.25M Tris-HCl, pH adjusted to 6.5

[00...

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Abstract

The invention discloses a magnetic bead method virus nucleic acid extraction kit and a use method, the magnetic bead method virus nucleic acid extraction kit comprises a lysis solution, a magnetic bead solution, a washing solution I, a washing solution II, a washing solution III and an eluent, the lysis solution is composed of the following substances: Tris-HCl, guanidine thiocyanate, EDTA, Triton-100, PEG-6000, SDS, isopropanol and water. The lysate provided by the invention has the beneficial effects that protease K does not need to be used, so that the cost is saved, heating lysis is generally needed during sample lysis, volatilization of an organic solvent can be accelerated, and the health of operators is harmed, but the lysate does not need to be heated, so that the harm to the operators can be reduced. The formula of the lysate for extracting nucleic acid is obtained by researching the proportion of the components of the lysate by the inventor.

Description

technical field [0001] The invention relates to the medical field, in particular to a magnetic bead virus nucleic acid extraction kit and a use method. Background technique [0002] In recent years, with the rapid development of molecular biology, as PCR and PR-PCR are more and more widely used in virus detection, the workload of nucleic acid extraction has increased rapidly. The traditional method of extracting nucleic acid is cumbersome and needs to be extracted by phenol or chloroform. , takes a long time and is highly toxic, and does not support automated extraction, which makes it unable to meet the requirements of large-scale disease detection. [0003] Nucleic acid extraction technologies that are simpler and more convenient to operate than traditional nucleic acid extraction methods have also appeared on the market, mainly including: chromatography column method and magnetic bead method. Since the chromatography column nucleic acid extraction technology requires the...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013C12N15/1003
Inventor 傅明华李美琼夏姝瑞聂晶
Owner 江苏迅睿生物技术有限公司
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