CN219117433U - Nucleic acid extraction tube, nucleic acid extraction kit and nucleic acid extraction system - Google Patents

Abstract

The utility model relates to the technical field of molecular biology, and discloses a nucleic acid extraction tube, a nucleic acid extraction kit and a nucleic acid extraction system, wherein the nucleic acid extraction tube comprises: the pipe body is provided with an upper layer sieve plate and a bottom layer sieve plate which are arranged in the pipe body; the upper end of the tube body is provided with a positive pressure port and a negative pressure port, and the lower end of the tube body is provided with an extraction reagent passing port; the cavity of the tube body between the upper layer sieve plate and the bottom layer sieve plate forms a nucleic acid extraction cavity, and solid-phase nucleic acid extraction resin is arranged in the nucleic acid extraction cavity. Based on the nucleic acid extraction tube and the corresponding nucleic acid extraction method, the problems of low extraction efficiency and easy pollution caused by the open space in the operation process in the prior art can be solved, and the pollution between samples in the nucleic acid extraction process is avoided; and the extraction process of the nucleic acid can be completed within 2-5 minutes from 30 minutes, so that the extraction efficiency of the nucleic acid is greatly improved.

Description

Nucleic acid extraction tube, nucleic acid extraction kit and nucleic acid extraction system

Technical Field

The utility model relates to the technical field of molecular biology, in particular to a nucleic acid extraction tube, a nucleic acid extraction kit and a nucleic acid extraction system.

Background

The molecular diagnosis process comprises the extraction of nucleic acid and the amplification detection of nucleic acid, and the nucleic acid amplification and detection based on temperature change or the isothermal nucleic acid amplification and detection provides a nucleic acid detection means with single-molecule level sensitivity. Among them, the extraction means of nucleic acid is a key factor for the effective detection of nucleic acid.

Common methods for extracting nucleic acid include a nucleic acid extraction method based on magnetic microspheres and a nucleic acid extraction method based on centrifugal columns (for example, a method for extracting genomic deoxyribonucleic acid, a centrifugal column for extracting nucleic acid, a nucleic acid extraction device and a nucleic acid extraction method of CN114657054a, which are disclosed in chinese published patent application CN113755487 a), wherein the two methods need to use corresponding necessary equipment such as a magnetic field and a centrifuge, and the nucleic acid extraction process involves a plurality of different treatments or multiple centrifuges; the time for nucleic acid extraction by these two methods is usually about 30 minutes. Therefore, there is a need in the art for a nucleic acid extraction system and a corresponding extraction structure to simplify the nucleic acid extraction process, so that the extraction process is simple and the extraction efficiency is high.

Disclosure of Invention

The utility model aims to overcome at least one defect of the prior art and provides a nucleic acid extraction tube, a nucleic acid extraction kit and a nucleic acid extraction system, so that the extraction process is simple and the extraction efficiency is high.

The technical scheme adopted by the utility model is that the nucleic acid extraction tube comprises:

the pipe body is provided with an upper layer sieve plate and a bottom layer sieve plate which are arranged in the pipe body; the upper end of the tube body is provided with a positive pressure port and a negative pressure port, and the lower end of the tube body is provided with an extraction reagent passing port; the body cavity of the upper layer sieve plate is provided with a nucleic acid extracting solution cavity; the cavity of the tube body between the upper layer sieve plate and the bottom layer sieve plate forms a nucleic acid extraction cavity, and solid-phase nucleic acid extraction resin is arranged in the nucleic acid extraction cavity.

The positive and negative pressure ports are at least used for communicating with the positive and negative pressure channels and the positive and negative pressure devices, and the extraction reagent passing ports are at least used for communicating with the nucleic acid extraction reagent supply container and the pipeline. Further, the positive and negative pressure ports are communicated with a positive and negative pressure channel and a positive and negative pressure device which are communicated with the sterile space. The positive and negative pressure ports of the nucleic acid extraction tube are communicated with a structure for providing positive and negative pressure, and the structure for providing positive and negative pressure comprises a pipette. In the process of extracting nucleic acid, the positive pressure port and the negative pressure port are kept in a closed state relative to the external environment, so that the operation in a completely opened state is avoided, pollutants in the air can be effectively reduced from falling into the nucleic acid extracting tube, and the pollution is reduced. Further, the extraction reagent passage port may be in communication with a conduit for transporting the nucleic acid extraction reagent or may be positioned below the level of the nucleic acid extraction reagent, reducing contamination during addition of the extraction reagent.

In the process, the solution to be extracted contacts with solid-phase nucleic acid extraction resin particles in the nucleic acid extraction cavity through a bottom layer sieve plate, and then enters the nucleic acid extraction liquid cavity through an upper layer sieve plate; in the discharging process, the solution to be extracted is discharged through an upper layer sieve plate, solid phase nucleic acid extraction resin and a bottom layer sieve plate; in the process of entering and discharging, the liquid to be extracted is driven by positive and negative pressure, so that the liquid to be extracted does not bypass the surface of part of the solid-phase nucleic acid extraction resin due to solid blockage, but can basically pass through all the solid-phase nucleic acid extraction resin smoothly, thereby realizing full contact; by the method, the adsorption of nucleic acid can be rapidly promoted, the combination effect is improved, the number of times of repeated positive and negative pressure is reduced, and the extraction efficiency of the resin adsorption process is improved. Meanwhile, in the positive and negative pressure process, the bottom layer sieve plate and the upper layer sieve plate can effectively screen out macromolecular substances, so that the macromolecular substances are prevented from being concentrated in the nucleic acid extraction cavity, the adsorption of solid-phase nucleic acid extraction resin on macromolecular impurities is reduced, the extraction rate is improved, and meanwhile, the binding sites occupied by the impurities on the surface of the resin can be reduced, so that the resin can be conveniently and rapidly bound with nucleic acid which can pass through the upper layer sieve plate and the bottom layer sieve plate. The extraction rate is improved, and simultaneously, the suction and discharge times of positive and negative pressure in the process are reduced, so that the extraction efficiency is further improved. In addition, the extraction rate in the process is improved, and the repeated operation times of the subsequent washing process are reduced conveniently. Similarly, in the subsequent washing process, the bottom screen plate and the upper screen plate also play a role in reducing the influence of impurities.

Further, the tube is formed as a pipette tube. The pipette body is manufactured, so that the pipette body is convenient for simplifying the manufacturing process of the existing equipment, is also convenient for being matched with a pipette of the prior art, and is simple to operate. The pipette may be a pipette tip mated with a pipette gun.

Further, the device also comprises an aerosol sieve plate arranged above the upper sieve plate in the tube body, and a nucleic acid extracting solution cavity is formed between the aerosol sieve plate and the upper sieve plate. By the aerosol sieve plate, air aerosol pollution can be further prevented, and sample pollution in the nucleic acid extraction process is reduced.

Further, the aperture of the aerosol sieve plate is 10-100 μm.

Further, the solid phase nucleic acid extraction resin is filled in a part of the nucleic acid extraction chamber or in the nucleic acid extraction chamber.

Further, the surface modification functional groups of the solid phase nucleic acid extraction resin include hydroxyl groups, silylhydroxyl groups, amine groups, carboxyl groups, and/or aldehyde groups. The surface modification functional group is beneficial to the rapid combination of nucleic acid molecules, effectively adsorbs nucleic acid and improves the nucleic acid extraction efficiency.

Further, the diameter of the solid phase nucleic acid extraction resin particles is 2 μm to 2mm.

Further, the aperture of the bottom layer sieve plate is 1 mu m-1 mm; and/or the aperture of the upper layer sieve plate is 1 mu m-1 mm.

It is another object of the present utility model to provide a nucleic acid extraction kit comprising the aforementioned nucleic acid extraction tube, and a lysate, an adsorbent, a wash solution, and/or an eluent. Further, the method comprises the step of extracting the nucleic acid from the nucleic acid extraction tube, and a lysate, an adsorption solution, a washing solution and an eluent.

It is still another object of the present utility model to provide a nucleic acid extraction method, based on any of the above nucleic acid extraction tubes, comprising the steps of:

s1, mixing a nucleic acid sample with a lysate to release nucleic acid in cells to the outside of the cells, and mixing an adsorption solution;

s2, enabling the solution obtained in the S1 to enter a nucleic acid extraction tube from a bottom layer sieve plate, contact with solid-phase nucleic acid extraction resin, and enter a nucleic acid extraction liquid cavity after passing through the solid-phase nucleic acid extraction resin and an upper layer sieve plate; then, the solution is discharged from the nucleic acid extracting solution cavity from the upper screen plate, and is discharged from the nucleic acid extracting tube through the solid-phase nucleic acid extracting resin and the bottom screen plate; repeatedly entering and discharging the nucleic acid extraction tube for a plurality of times, and finally discharging the solution from the nucleic acid extraction tube;

s3, sucking the washing liquid into a nucleic acid extraction tube from a bottom layer sieve plate, contacting with the solid-phase nucleic acid extraction resin, and allowing the washing liquid to enter a nucleic acid extraction liquid cavity through the solid-phase nucleic acid extraction resin and an upper layer sieve plate; then, the washing liquid is discharged from the nucleic acid extracting liquid cavity from the upper layer sieve plate, and is discharged from the nucleic acid extracting tube through the solid-phase nucleic acid extracting resin and the bottom layer sieve plate; repeatedly entering and discharging the nucleic acid extraction tube for a plurality of times, and finally discharging the solution from the nucleic acid extraction tube;

s4, enabling the eluent to enter a nucleic acid extraction tube, and discharging the eluent out of the nucleic acid extraction tube after elution to finish nucleic acid extraction.

Further, in the step S2, the solution obtained in the step S1 is sucked into a nucleic acid extraction tube from an extraction reagent through a port and a bottom screen plate by negative pressure, and the solution S1 contacts with solid-phase nucleic acid extraction resin and enters a nucleic acid extraction liquid cavity through the solid-phase nucleic acid extraction resin and an upper screen plate; then, positive pressure is carried out again to enable the solution to be discharged from the nucleic acid extracting solution cavity from the upper layer sieve plate, and the solution is discharged from the nucleic acid extracting tube through the solid-phase nucleic acid extracting resin and the bottom layer sieve plate from the extracting reagent through hole; repeatedly sucking and discharging the nucleic acid extraction tube for multiple times, and finally discharging the solution out of the nucleic acid extraction tube;

further, in step S3, negative pressure causes the washing liquid to be sucked into the nucleic acid extraction tube from the extraction reagent passing port and the bottom sieve plate, and causes the washing liquid to contact with the solid-phase nucleic acid extraction resin and enter the nucleic acid extraction liquid cavity through the solid-phase nucleic acid extraction resin and the upper sieve plate; then positive pressure is used for discharging the washing liquid from the nucleic acid extracting liquid cavity from the upper layer sieve plate, and the washing liquid is discharged from the nucleic acid extracting pipe through the solid phase nucleic acid extracting resin and the bottom layer sieve plate from the extracting reagent through hole; repeatedly sucking and discharging the nucleic acid extraction tube for a plurality of times, and finally discharging all the washing liquid out of the nucleic acid extraction tube;

further, in step S4, the eluent is sucked into the nucleic acid extraction tube through the bottom layer sieve plate by negative pressure, contacted with the solid phase nucleic acid extraction resin for several seconds to several minutes, and then discharged out of the nucleic acid extraction tube by positive pressure.

It is still another object of the present utility model to provide a nucleic acid extraction system comprising the aforementioned nucleic acid extraction tube, and positive and negative pressure driving means. Further, the positive and negative pressure driving mechanism is a pipette gun.

Compared with the prior art, the utility model has the beneficial effects that: the nucleic acid extraction tube and the nucleic acid extraction method based on the same solve the problems of low extraction efficiency and easy pollution in an open space in the operation process in the prior art, and the whole process of extracting the nucleic acid is completed in a relatively closed nucleic acid extraction tube, including adsorption, washing and elution of the nucleic acid, so that the pollution among samples in the process of extracting the nucleic acid is avoided; in addition, in the nucleic acid extraction process, the elution of the nucleic acid does not need heating, so that the pollution between samples caused by aerosol generated in the heating process can be effectively avoided. Through the scheme of the application, the pollution in the process of extracting the nucleic acid can be effectively reduced from multiple aspects, and the effective extraction of the nucleic acid is facilitated. Meanwhile, through the nucleic acid extraction tube and the extraction method provided by the application, the nucleic acid extraction efficiency is also obviously improved, the extraction process of nucleic acid can be shortened from about 30 minutes required by the traditional nucleic acid extraction process to only 2-5 minutes, and the nucleic acid extraction steps are greatly simplified.

Drawings

Fig. 1 is a schematic structural diagram (one) of the present utility model.

Fig. 2 is a schematic structural diagram (ii) of the present utility model.

Description of the drawings: 100. a nucleic acid extraction tube; 101. a positive and negative pressure port; 102. an aerosol screen plate; 103. a nucleic acid extraction liquid chamber; 104. an upper layer sieve plate; 105. a nucleic acid extraction chamber; 106. a solid phase nucleic acid extraction resin; 107. a bottom layer sieve plate; 108. the extraction reagent passes through the port.

Detailed Description

The drawings are for illustrative purposes only and are not to be construed as limiting the utility model. For better illustration of the following embodiments, some parts of the drawings may be omitted, enlarged or reduced, and do not represent the actual product dimensions; it will be appreciated by those skilled in the art that certain well-known structures in the drawings and descriptions thereof may be omitted.

The present embodiment provides a nucleic acid extraction cuvette comprising: a tube body, an upper layer sieve plate 104 and a bottom layer sieve plate 107 which are arranged in the tube body; the upper end of the tube body is provided with a positive pressure port and a negative pressure port 101, the lower end of the tube body is provided with an extraction reagent passing port 108, and the positive pressure port and the negative pressure port 101 are communicated with the extraction reagent passing port 108; the body cavity of the upper layer sieve plate 104 is provided with a nucleic acid extracting solution cavity 103; the cavity of the tube body between the upper layer sieve plate 104 and the bottom layer sieve plate forms a nucleic acid extraction cavity 105, a solid phase nucleic acid extraction resin 106 which is partially filled or a solid phase nucleic acid extraction resin 106 which is filled is arranged in the nucleic acid extraction cavity 105, the diameter of particles of the solid phase nucleic acid extraction resin 106 is 2 mu m-2 mm, and the surface modification functional groups of the solid phase nucleic acid extraction resin 106 comprise hydroxyl, silicon hydroxyl, amino, carboxyl and/or aldehyde groups. In order to further reduce the possibility of the influence of aerosol, an aerosol screen plate 102 may be disposed above the upper screen plate 104 in the tube, the aperture of the aerosol screen plate 102 is 10 μm to 100 μm, the influence of aerosol is further prevented by the aerosol screen plate 102, and when the aerosol screen plate 102 is disposed, the nucleic acid extraction liquid cavity 103 is formed by the space between the aerosol screen plate 102 and the upper screen plate 104.

The aperture of the bottom layer sieve plate is 1 mu m-1 mm; and/or the aperture of the upper layer screen plate 104 is 1 μm-1 mm.

The tube body of the nucleic acid extraction tube 100 in this embodiment may also be formed as a pipette. The pipette formation specifically comprises the following manufacturing processes: taking a pipette, placing a sieve plate with the aperture of 1 μm-1 mm at the bottom of the pipette to form a bottom sieve plate 107, filling part of the nucleic acid extraction cavity 105 with the solid-phase nucleic acid extraction resin 106 or filling the nucleic acid extraction cavity 105, placing the sieve plate with the aperture of 1 μm-1 mm in the pipette to form an upper sieve plate 104, and forming a nucleic acid extraction liquid cavity 103 above the upper sieve plate 104 of the pipette body to complete the preparation of the nucleic acid extraction tube 100. The upper opening of the pipette tube body serves as a positive and negative pressure port 101, and the lower opening of the pipette serves as an extraction reagent passing port 108. Furthermore, after the upper screen plate 104 is placed, a screen plate having a pore diameter of 10 μm to 100 μm may be placed over the upper screen plate 104 to form an aerosol screen plate 102, and a nucleic acid extraction solution chamber 103 may be formed between the aerosol screen plate 102 and the upper screen plate 104. The structure is shown in fig. 1.

The present embodiment also provides a nucleic acid extraction kit comprising the aforementioned nucleic acid extraction tube 100, and a lysis solution, an adsorption solution, a washing solution, and/or an eluent.

The embodiment also provides a nucleic acid extraction method, which is performed based on the nucleic acid extraction tube, and comprises the following steps:

s1, mixing a nucleic acid sample with a lysate to release nucleic acid in cells to the outside of the cells, and mixing an adsorption solution;

s2, enabling the solution obtained in the S1 to enter a nucleic acid extraction tube 100 from a bottom layer sieve plate 107, contact with a solid-phase nucleic acid extraction resin 106, and enter a nucleic acid extraction liquid cavity after passing through the solid-phase nucleic acid extraction resin 106 and an upper layer sieve plate 104; then, the solution is discharged from the nucleic acid extraction solution chamber from the upper sieve plate, and discharged from the nucleic acid extraction tube 100 through the solid phase nucleic acid extraction resin 106 and the bottom sieve plate 107; repeatedly entering and exiting the nucleic acid extraction tube 100 a plurality of times, and finally allowing the solution to be completely discharged from the nucleic acid extraction tube 100;

s3, sucking the washing liquid into the nucleic acid extraction tube 100 from the bottom layer sieve plate 107, contacting with the solid phase nucleic acid extraction resin 106, and entering a nucleic acid extraction liquid cavity through the solid phase nucleic acid extraction resin 106 and the upper layer sieve plate 104; then, the washing solution is discharged from the nucleic acid extraction solution chamber from the upper screen plate, and discharged from the nucleic acid extraction tube 100 through the solid phase nucleic acid extraction resin 106 and the bottom screen plate 107; repeatedly entering and exiting the nucleic acid extraction tube 100 a plurality of times, and finally allowing the solution to be completely discharged from the nucleic acid extraction tube 100;

s4, enabling the eluent to enter the nucleic acid extraction tube 100, and discharging the eluent out of the nucleic acid extraction tube 100 after elution to finish nucleic acid extraction.

Example 1

The body of the nucleic acid extraction tube 100 in this example was formed as a 200. Mu.l pipette. The preparation method specifically comprises the following nucleic acid extraction tube preparation processes: taking a 200ul pipette, placing a sieve plate with the diameter of 1mm and the aperture of 50 micrometers at the bottom of the pipette to form a bottom sieve plate 107, filling part of the nucleic acid extraction cavity 105 with 5mg of microcrystalline cellulose serving as solid-phase nucleic acid extraction resin 106, placing a sieve plate with the diameter of 3mm and the aperture of 50 micrometers in the pipette to form an upper sieve plate 104, and forming a nucleic acid extraction liquid cavity 103 above the upper sieve plate of the pipette body to complete the preparation of the nucleic acid extraction tube 100. The upper opening of the pipette tube body serves as a positive and negative pressure port 101, and the lower opening of the pipette serves as an extraction reagent passing port 108. The structure is shown in fig. 2. In addition to the structure of fig. 2, an aerosol screen panel 102 may be added over the upper screen panel 104 to further reduce contamination during nucleic acid extraction.

After completion of the preparation of the nucleic acid extraction tube, the present example performed a nucleic acid extraction operation through the nucleic acid extraction tube 100, and 20. Mu.l of the whole blood sample was mixed and lysed with a proteinase K-containing lysate, and then further mixed with a 5M NaCl adsorption solution containing 30% PEG-6000.

The nucleic acid extraction tube 100 is placed on a pipette, and in a specific operation, the positive and negative pressure port 101 of the nucleic acid extraction tube is connected with the tip of the pipette. Then, the solution is sucked into the nucleic acid extraction tube 100 from the extraction reagent passing port 108 and the bottom layer sieve plate 107 by using the negative pressure of the pipette and is brought into contact with the solid phase nucleic acid extraction resin 106, and then the solution is discharged out of the nucleic acid extraction tube 100 by using the positive pressure, and the negative pressure suction and positive pressure discharge processes are repeated for 2 times, and finally the solution is discharged out of the nucleic acid extraction tube 100. In the suction process, the solution enters the nucleic acid extraction cavity 105 positioned at the other side of the bottom screen plate 107 from one side of the bottom screen plate and contacts with the solid-phase nucleic acid extraction resin, so that the nucleic acid is adsorbed on the solid-phase nucleic acid extraction resin, then enters the nucleic acid extraction liquid cavity through the upper screen plate, then is discharged at positive pressure, and contacts with the solid-phase nucleic acid extraction resin again when passing through the nucleic acid extraction cavity 105, and finally the solution is discharged through the bottom screen plate 107. In the single negative pressure suction and positive pressure discharge processes, the bottom layer sieve plate 107 and the upper layer sieve plate play a role in removing macromolecular impurities, so that small molecular substances such as nucleic acid and the like are concentrated in the nucleic acid extraction cavity 105, impurities adsorbed by the solid-phase nucleic acid extraction resin are reduced, and the extraction efficiency is promoted. The method can realize that the nucleic acid in the solution is basically adsorbed by the solid-phase nucleic acid extraction resin by repeating for a plurality of times, and avoid the solid-phase nucleic acid extraction resin from adsorbing macromolecules and impurities which are not easy to remove. In the process, the positive and negative pressure drives the solution to be sucked and discharged, and the solution is completely contacted with the solid-phase nucleic acid extraction resin in the process of sucking and discharging the solution from the nucleic acid extraction tube, so that the nucleic acid extraction operation is simplified, and the extraction efficiency is effectively improved.

Then, the nucleic acid-extracted tube was aspirated with a washing solution containing 1M KCl, and washed 2-3 times. Washing is carried out in the same process of sucking and discharging the nucleic acid-containing solution, so that the washing liquid is convenient to contact with the solid-phase nucleic acid extraction resin to wash away adsorbed impurities, and the interference of the impurities is reduced.

Finally, the DNA was eluted out of the nucleic acid eluting tube with 25ul of 10mM Tris (ph=8) eluate. Eluting the nucleic acid adsorbed on the solid phase nucleic acid extraction resin by using an eluent to obtain the extracted nucleic acid.

In this example, a 260/a280=1.9 of DNA in the eluate after elution, and the nucleic acid extraction efficiency was about 70%. Through the nucleic acid extraction tube and the corresponding nucleic acid extraction method, nucleic acid can be effectively extracted, the process is simple, and the extraction efficiency is high.

Example 2

Similar to example 1, but in this example, the nucleic acid extraction tube 100 was formed as a 1ml pipette. The preparation method specifically comprises the following nucleic acid extraction tube preparation processes: placing a sieve plate with the diameter of 3mm and the aperture of 10 micrometers at the bottom of a pipette to form a bottom sieve plate 107, filling a part of nucleic acid extraction cavity 105 by using 15mg of Diethylaminoethyl (DEAE) modified 100 micrometer microcrystalline cellulose as solid-phase nucleic acid extraction resin 106, placing a sieve plate with the diameter of 6mm and the aperture of 10 micrometers in the pipette to form an upper sieve plate 104, forming a nucleating acid extraction liquid cavity 103 above the upper sieve plate of a pipette body, and completing the manufacture of the nucleic acid extraction tube 100. The upper opening of the pipette tube body serves as a positive and negative pressure port 101, and the lower opening of the pipette serves as an extraction reagent passing port 108. The structure is shown in fig. 2.

After completion of the preparation of the nucleic acid extraction tube, the present example performs a nucleic acid extraction operation through the nucleic acid extraction tube 100, mixes 200. Mu.l of the liquefied sputum with 200. Mu.l of a lysate containing proteinase K and 2M guanidinium isothiocyanate and heats it for 10 minutes at 50℃and then for 5 minutes at 95℃and mixes the solution with 200. Mu.l of an adsorption solution of 0.1M Tris (pH=7.5);

the positive and negative pressure ports 101 of the nucleic acid extraction tube are connected to a mechanism for supplying positive and negative pressure. Then, the solution is sucked into the nucleic acid extraction tube 100 from the extraction reagent passing port 108 and the bottom layer sieve plate 107 by the negative pressure and is brought into contact with the solid phase nucleic acid extraction resin 106, and then the solution is discharged out of the nucleic acid extraction tube 100 by the positive pressure, and the negative pressure suction and positive pressure discharge processes are repeated 3 times, and finally the solution is discharged out of the nucleic acid extraction tube 100.

Then, 10mM Tris (pH=8) and 60% ethanol were used as washing solutions to draw the nucleic acid extraction tube 100 into the nucleic acid extraction tube, and washing was performed 2 to 3 times. The operation is the same as the solution suction and discharge process described above, except that the suction and discharge of the washing solution is replaced.

Finally, RNA was eluted out of the nucleic acid eluting tube with 50ul of 2mM NaOH (pH=11) containing 0.3M KCl as eluent. Eluting the nucleic acid adsorbed on the solid phase nucleic acid extraction resin by using an eluent to obtain the extracted nucleic acid.

In this example, 2ul of the eluate was added to the PCR solution for real-time PCR, and the Ct value of RNA showed that the extraction efficiency of RNA was not less than 10%. Namely, by the nucleic acid extraction tube and the nucleic acid extraction method provided by the embodiment, the effective extraction of nucleic acid is realized as well.

Example 3

Similar to example 1, but in this example, the nucleic acid extraction tube 100 was formed as a 2ml pipette. The preparation method specifically comprises the following nucleic acid extraction tube preparation processes: taking a 2ml pipette, placing a sieve plate with the diameter of 3mm and the aperture of 10 micrometers at the bottom of the pipette to form a bottom sieve plate 107, taking 10mg of 5 micrometer glass beads as solid-phase nucleic acid extraction resin 106 to fill a nucleic acid extraction cavity 103, placing a sieve plate with the diameter of 4mm and the aperture of 10 micrometers in the pipette to form an upper sieve plate 104, and forming a nucleic acid extraction liquid cavity 103 above the upper sieve plate of the pipette body to complete the preparation of the nucleic acid extraction tube 100. The upper opening of the pipette tube body serves as a positive and negative pressure port 101, and the lower opening of the pipette serves as an extraction reagent passing port 108. The structure is shown in fig. 2.

After completion of the preparation of the nucleic acid extraction tube, the present example performs a nucleic acid extraction operation through the nucleic acid extraction tube 100, mixes 100 microliters of throat swab sample with 200 microliters of lysate containing proteinase k and 4M guanidinium isothiocyanate and heats at 60 degrees celsius for 10 minutes, then mixes the solution with 100 microliters of adsorption solution of 0.1M citric acid (ph=4);

the positive and negative pressure ports 101 of the nucleic acid extraction tube are connected to a mechanism for supplying positive and negative pressure. Then, the solution is sucked into the nucleic acid extraction tube 100 from the extraction reagent passing port 108 and the bottom layer sieve plate 107 by negative pressure and passed through the nucleic acid extraction resin 106, and then the solution is discharged out of the nucleic acid extraction tube 100 by positive pressure, and this negative pressure suction and positive pressure discharge process is repeated 2 times, and finally the solution is discharged out of the nucleic acid extraction tube 100.

Then, 60% ethanol was used as a washing solution to be sucked into the nucleic acid extraction tube 100 and washed 2 to 3 times. The operation is the same as the solution suction and discharge process described above, except that the suction and discharge of the washing solution is replaced.

Finally, the DNA was eluted out of the nucleic acid eluting tube with 100ul of 10mM Tris (ph=9) as eluent. Eluting the nucleic acid adsorbed on the solid phase nucleic acid extraction resin by using an eluent to obtain the extracted nucleic acid.

In this example, 5ul of the eluate was added to the PCR solution for the real-time PCR, and the Ct value of the DNA showed that the extraction efficiency of the DNA was not less than 40%. By combining the previous embodiments, the technical scheme provided by the application can have an effective extraction effect on DNA and RNA nucleic acid. And the extraction process is simple and high in efficiency, and in the extraction process of the embodiment 1-3, the complete extraction time is only 2-5 min. Meanwhile, the nucleic acid extraction tube and the extraction method based on the improvement of the application also avoid heating in the elution process and reduce the pollution of aerosol in the heating process.

Example 4

The present embodiment provides a nucleic acid extraction system, which includes the nucleic acid extraction tube 100 in the foregoing implementation process, and a positive and negative pressure driving mechanism; the positive and negative pressure driving mechanism is connected with the positive and negative pressure port 101 of the nucleic acid extracting tube and provides positive and negative pressure for the positive and negative pressure port 101. The positive and negative pressure driving mechanism can be a manual pipette or an electric pipette.

It should be understood that the foregoing examples of the present utility model are merely illustrative of the present utility model and are not intended to limit the present utility model to the specific embodiments thereof. Any modification, equivalent replacement, improvement, etc. that comes within the spirit and principle of the claims of the present utility model should be included in the protection scope of the claims of the present utility model.

Claims (10)

1. A nucleic acid extraction tube, comprising:

the pipe body is provided with an upper layer sieve plate and a bottom layer sieve plate which are arranged in the pipe body; the upper end of the tube body is provided with a positive pressure port and a negative pressure port, and the lower end of the tube body is provided with an extraction reagent passing port; the cavity of the tube body between the upper layer sieve plate and the bottom layer sieve plate forms a nucleic acid extraction cavity, and solid-phase nucleic acid extraction resin is arranged in the nucleic acid extraction cavity.

2. The nucleic acid extraction tube of claim 1, wherein the tube body is a pipette tube body.

3. The nucleic acid extraction tube of claim 1, further comprising an aerosol screen panel disposed within the tube above the upper screen panel, wherein a nucleic acid extraction liquid chamber is formed between the aerosol screen panel and the upper screen panel.

4. The nucleic acid extraction tube according to claim 3, wherein the aerosol screen plate has a pore size of 10 μm to 100. Mu.m.

5. The nucleic acid extraction tube according to any one of claims 1 to 4, wherein the solid phase nucleic acid extraction resin fills the nucleic acid extraction chamber partially or fully.

6. The nucleic acid extraction tube according to any one of claims 1 to 4, wherein the surface modification functional group of the solid phase nucleic acid extraction resin comprises a hydroxyl group, a silylhydroxyl group, an amine group, a carboxyl group and/or an aldehyde group.

7. The nucleic acid extraction tube according to any one of claims 1 to 4, wherein the diameter of the solid phase nucleic acid extraction resin particles is 2 μm to 2mm.

8. The nucleic acid extraction tube according to any one of claims 1 to 4, wherein the pore diameter of the bottom layer sieve plate is 1 μm to 1mm; and/or the aperture of the upper layer sieve plate is 1 mu m-1 mm.

9. A nucleic acid extraction kit comprising the nucleic acid extraction tube of any one of claims 1 to 8, and a lysate, an adsorbent, a wash solution, and/or an eluent.

10. A nucleic acid extraction system comprising the nucleic acid extraction tube of any one of claims 1 to 8, and a positive and negative pressure driving mechanism; and/or the positive and negative pressure driving mechanism is a manual pipette or an electric pipette.

Images