Rapid nucleic acid extraction device and method

An extraction device and extraction method technology, applied in the direction of sterilization methods, biochemical cleaning devices, biochemical equipment and methods, etc., can solve the problems of nucleic acid loss, cross-contamination, and liquid flying out, so as to improve extraction efficiency and avoid Effect of cross-contamination and improving nucleic acid extraction efficiency

Active Publication Date: 2015-06-10
奥健生物科技(广州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In addition, the steps of lysis, binding, and washing are carried out in different centrifuge tubes, and the solution containing nucleic acid needs to be tra

Method used

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  • Rapid nucleic acid extraction device and method
  • Rapid nucleic acid extraction device and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Whole blood DNA in the swab specimen of the rapid nucleic acid extraction device of the present invention comprises the following steps:

[0048] (1) Prepare simulated swab material

[0049] Put a sterile Omni swab into the first cavity 21 of the spin column of the present invention, press the end of the rod, eject the swab, and add 50 ul of fresh EDTA anticoagulated mouse whole blood to the swab as a simulated swab sample.

[0050] (2) cracking

[0051] Add 200ul lysis buffer (Lysis buffer) and 20ul proteinase K to the swab, cover the tube cap, and lyse at 65°C for 20min.

[0052] (3) combine

[0053] Add 200ul of binding buffer to the swab, mix well, and centrifuge at 20000g for 2min.

[0054] (4) rinse

[0055] Add 500ul of wash buffer A (wash buffer A) to the first cavity of the spin column, cover the cap, centrifuge at 12000g for 1min, transfer the spin column to a new collection tube, add 500ul of wash buffer B (wash buffer B), Cover the tube cap, centrifuge ...

Embodiment 2

[0058] The conventional spin column method for extracting whole blood DNA from swab samples includes the following steps:

[0059] (1) Prepare simulated swab material

[0060] Put the sterile Omni swab into a 1.5ml centrifuge tube, press the end of the rod, eject the swab, and add 50ul of fresh EDTA anticoagulated mouse whole blood to the swab as a simulated swab sample.

[0061] (2) cracking

[0062] Add 200ul Lysis buffer and 20ul proteinase K to the swab, cover the tube cap, and lyse at 65°C for 20min.

[0063] (3) combine

[0064] Add 200ul binding buffer to the swab, mix well, centrifuge at 12000g for 1min, transfer the supernatant to a conventional spin column, centrifuge at 12000g for 1min, and transfer the spin column to a new collection tube.

[0065] (4) rinse

[0066]Add 500ul wash buffer A to the spin column, cover the cap, centrifuge at 12000g for 1min, transfer the spin column to a new collection tube, add 500ul wash buffer B, cap the tube, centrifuge again a...

Embodiment 3

[0069] The nucleic acid rapid extraction device of the present invention extracts DNA in blood spot samples, comprising the following steps:

[0070] (1) Prepare simulated blood spot samples

[0071] Cut off 0.5 cm 2 For denim fabric, add 50ul of fresh EDTA anticoagulated mouse whole blood to the denim fabric, leave it at room temperature for 1 hour, let it dry naturally, and use it as a simulated blood spot specimen. Then cut the simulated blood spot sample into small pieces and put them into the first cavity 21 of the spin column.

[0072] (2) cracking

[0073] Add 200ul Lysis buffer and 20ul proteinase K to the simulated blood spot sample, cover the tube cap, and lyse at 65°C for 20min.

[0074] (3) combine

[0075] Add 200ul binding buffer to the simulated blood spot, mix well, and centrifuge at 20000g for 2min.

[0076] (4) rinse

[0077] Add 500ul wash buffer A to the first chamber 21 of the spin column, cover the cap, centrifuge at 12000g for 1min, transfer the sp...

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Abstract

The invention provides a rapid nucleic acid extraction device which comprises a centrifugal column, wherein the centrifugal column comprises two independent cavities formed by separation; the independent cavities comprise a first cavity and a second cavity; a vertical separator and a horizontal separator through which two independent cavities are formed are arranged between the first cavity and the second cavity; a micro-porous filtration membrane and a nucleic acid adsorption membrane are arranged in the centrifugal column; an opening which is respectively connected with the first cavity and the second cavity is formed in the top of the centrifugal column; and the nucleic acid adsorption membrane is arranged on the inner side at the bottom of the centrifugal column. The invention also provides a rapid nucleic acid extraction method realized based on the rapid nucleic acid extraction device. According to the technical scheme disclosed by the invention, aiming at the nucleic acid extraction process of trace samples (such as medicolegal expertise), losses of trace nucleic acid material evidences caused by adsorption of checked materials such as swabs, cotton swabs, seminal stains, blood cakes and cigarette ends and adsorption of the centrifugal tube wall can be effectively avoided, the nucleic acid extraction efficiency is improved, and cross contamination is reduced.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a rapid nucleic acid extraction device for laboratory or medical testing and a rapid nucleic acid extraction method based on the rapid nucleic acid extraction device. Background technique [0002] At present, in the field of biotechnology, the traditional spin column method or magnetic bead method is used for nucleic acid extraction, which generally requires four steps of lysis, binding, rinsing, and elution. First, proteinase K, surfactant and other components are used to destroy the cell structure and release the nucleic acid into the solution; then, add high-concentration chaotropic salt and ethanol and other components to combine the nucleic acid with the adsorption membrane or magnetic beads; then the adsorption membrane or The magnetic beads are washed to remove inhibitors; finally, the nucleic acids on the adsorption membrane or magnetic beads are eluted. In the process of fo...

Claims

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Application Information

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IPC IPC(8): C12M1/24C12M1/12C12N15/10
CPCC12N15/10
Inventor 余家昌杜正平熊槐
Owner 奥健生物科技(广州)有限公司
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