A combined magnetic bead reaction tube and method of use thereof

Abstract

The application discloses a combined magnetic bead reaction tube and a using method thereof. The first plate body is sleeved above the outer wall of the lower tube body and extends to one side of the lower tube body in the horizontal direction. The outer side of the extension section and one end of the second plate body are arranged in an open and close mode, and the outermost upper end and the lower side of the other end of the second plate body are in contact. The lower tube cover is arranged at the position corresponding to the opening of the lower tube body, and the outer diameter is equal to the inner diameter of the opening of the lower tube body. The upper tube body corresponding to the position of the lower tube cover is fixed on the second plate body. The lower end of the third plate body is fixedly connected with the other end of the second plate body, and the upper end of the third plate body is flush with the upper tube body. One end of the fourth plate body is arranged in an open and close mode with the upper end of the third plate body, and the side of the other end of the fourth plate body downward is fixed with the magnetic body which can be inserted into the upper tube body. The height of the magnetic body is the same as that of the upper tube body. In use, the magnetic beads can be released without opening the cover, and the function of adsorbing the magnetic beads is realized.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, and in particular to a combined magnetic bead reaction tube and its method of use. Background Technology

[0002] Magnetic beads, as a novel extraction reagent frequently used in reactions such as nucleic acid extraction, immunology, and molecular purification, have been widely used in immunoassay, immunodetection, cell separation, purification of biological macromolecules, and molecular biology since their invention in the 1990s.

[0003] Magnetic beads possess unique physicochemical properties, such as reducing reaction rates and affecting reaction results in reaction systems. Adding magnetic beads during experiments necessitates opening the container during the reaction process, which can contaminate the sample and potentially cause aerosol pollution, further hindering the reaction. For samples with biohazardous properties, the released aerosols pose a significant risk to the health and safety of operators. Summary of the Invention

[0004] To address the problems existing in the prior art, the present invention provides a combined magnetic bead reaction tube and its usage method, which can release magnetic beads without opening the cap and achieve the function of adsorbing magnetic beads.

[0005] This invention is achieved through the following technical solution:

[0006] A combined magnetic bead reaction tube includes a lower tube body, an upper tube body, a first connector, and a second connector.

[0007] The lower tube body is open. The first connecting member includes a first plate and a second plate. The first plate is sleeved on the upper part of the outer wall of the lower tube body. The first plate and the opening of the lower tube body are flush. The first plate extends horizontally to one side of the lower tube body. The second plate is a planar structure. The outer side of the extension of the first plate and one end of the second plate are open and closed. The upper outermost end of the extension of the first plate and the lower side of one end of the second plate are in contact. A cylindrical lower tube cover is provided on the second plate at the position corresponding to the opening of the lower tube body. The outer diameter of the lower tube cover is equal to the inner diameter of the opening of the lower tube body. The other end of the second plate is located on one side of the lower tube body.

[0008] The second plate is fixed with an upper tube body corresponding to the position of the lower tube cover. The second connector includes a third plate body and a fourth plate body. The lower end of the third plate body is fixedly connected to the other end of the second plate body. The upper end of the third plate body is away from the upper tube body and is flush with the upper tube body. One end of the fourth plate body is open and closed with the upper end of the third plate body. A magnetic body that can be inserted into the upper tube body is fixed on the downward side of the other end of the fourth plate body. The magnetic body is at the same height as the upper tube body.

[0009] Preferably, it also includes a cylindrical upper tube cap fixed to the lower side of the other end of the fourth plate. The height of the upper tube cap is equal to or less than that of the upper tube body. The magnetic body is a cylinder with an outer diameter equal to the inner diameter of the upper tube body. The magnetic body is fixedly inserted into the upper tube body.

[0010] Furthermore, the height of the upper tube cover is less than that of the upper tube body, and the lower end of the magnet is located below the upper tube cover.

[0011] Preferably, the upper tube cover, lower tube cover, lower tube body, upper tube body, first connector and second connector are all made of PC plastic.

[0012] Preferably, the magnetic material is a ferrite magnet, a rare earth magnet, an AlNiCo alloy magnet, or a polymer magnetic compound.

[0013] Preferably, the height of the upper tube is 3-8mm.

[0014] Preferably, the number of upper tubes and magnetic bodies is the same and there are several upper tubes, which are evenly arranged on the second plate.

[0015] Preferably, the first plate and the second plate are integral structures, and the third plate and the fourth plate are integral structures.

[0016] Preferably, the lower tube body comprises, from top to bottom, an integral cylindrical section, a transition section, and a frustum section. The outer diameter of the upper end of the transition section is the same as the outer diameter of the lower end of the cylindrical section, and the outer diameter of the lower end of the transition section is the same as the outer diameter of the upper end of the frustum section. The lower end of the lower tube body is hemispherical or conical, and the diameter of the bottom surface of the hemispherical or conical shape is equal to the diameter of the bottom surface of the frustum section. The inner diameter of the cylindrical section is the same as the outer diameter of the lower tube cover.

[0017] A method of using a combined magnetic bead reaction tube, based on any one of the combined magnetic bead reaction tubes described above, includes the following steps:

[0018] The fourth plate is pressed down, the magnetic body is inserted into the upper tube, and the corresponding magnetic beads are placed into the lower tube cover according to the reaction requirements. The magnetic body attracts the magnetic beads through magnetic force, and then the corresponding reaction is carried out.

[0019] When it is time to add magnetic beads, open the fourth plate and the magnetic beads enter the lower tube to participate in the reaction. After the reaction is complete, insert a magnetic body into the upper tube and invert the reaction tube. All the magnetic beads will be attracted into the lower tube cap.

[0020] Compared with the prior art, the present invention has the following beneficial technical effects:

[0021] This invention discloses a combined magnetic bead reaction tube. A cylindrical lower tube cap can be tightly inserted into the opening of the lower tube body. The first and second plates are joined together and in contact, ensuring the airtightness of the magnetic bead reaction. The other end of the second plate is located on one side of the lower tube body, allowing for easy opening of the lower tube cap after the reaction. A fourth plate, carrying a magnetic element, is inserted into the upper tube body and contacts the second plate. Magnetic force attracts the required magnetic beads to the lower tube cap. The magnetic element is then opened as needed, allowing the magnetic beads to participate in the reaction. After the reaction, the magnetic element is inserted into the upper tube body, and the combined magnetic bead reaction tube is inverted to attract all the magnetic beads, completing the reaction. This achieves the function of adding and recovering magnetic beads. This invention utilizes a magnetic element to control the release of magnetic beads, and the lower tube cap preserves the beads, avoiding the drawback of traditional reaction devices that require opening the cap midway to add magnetic beads. This effectively solves the problems of contamination and biological hazards caused by opening the cap during the reaction. Attached Figure Description

[0022] Figure 1 This is a schematic diagram of the magnetic bead reaction tube described in this invention from one perspective.

[0023] Figure 2 This is a schematic diagram of the magnetic bead reaction tube described in this invention from another perspective.

[0024] In the diagram: 1-Magnetic body, 2-Upper tube cap, 3-Lower tube cap, 4-Lower tube body, 5-First connector, 6-Upper tube body, 7-Second connector. Detailed Implementation

[0025] The present invention will be further described below with reference to the accompanying drawings and embodiments, but this should not be construed as limiting the present invention.

[0026] The present invention provides a combined magnetic bead reaction tube, comprising a magnetic body 1, an upper tube cap 2, a lower tube cap 3, a lower tube body 4, a first connector 5, an upper tube body 6, and a second connector 7. The first connector 5 comprises a first plate and a second plate, while the second connector 7 comprises a third plate and a fourth plate.

[0027] Specifically, such as Figure 1 and Figure 2As shown, on the one hand, the lower tube 4 is used to carry the reaction system and is designed with a common structure. Its upper end is open. The main structure of the first plate is a ring. The ring is fitted on the upper part of the outer wall of the lower tube 4 and is flush with the opening of the lower tube 4. The ring extends horizontally to one side of the lower tube 4. The extension section is rectangular. The second plate is a planar plate structure. The outer side of the extension section of the first plate and one end of the second plate are connected. At the same time, the uppermost outer end of the extension section contacts the lower side of one end of the second plate. In this way, the second plate can contact the first plate when it is closed. A lower tube cover 3 is provided at the position corresponding to the opening of the lower tube 4. It is cylindrical and the outer diameter is equal to the inner diameter of the opening of the lower tube 4. In this way, the lower tube cover 3 can be tightly inserted into the opening of the lower tube 4. The other end of the second plate is located on one side of the lower tube 4. In this way, the second plate extends outside the lower tube 4 and is designed with a triangular protrusion to facilitate opening the lower tube cover 3.

[0028] On the other hand, the magnetic beads can be of many types according to the needs of the magnetic bead reaction. Therefore, several upper tubes 6 corresponding to the position of the lower tube cover 3 are fixed on the second plate to fix magnetic beads of different specifications. The lower tube cover 3 stores magnetic beads of various specifications. The lower end of the third plate is perpendicular to the upper surface of the second plate, and the lower end of the third plate is fixedly connected to the other end of the second plate. The third plate is away from the upper tube 6 and is flush with the upper tube 6. One end of the fourth plate is open and closed with the upper end of the third plate. The other end of the fourth plate is fixed with a magnetic body 1 that can be inserted into the upper tube 6. The number of upper tubes 6 and magnetic bodies 1 is the same and there are several. Several upper tubes 6 are spaced apart and evenly arranged on the second plate. At the same time, it is necessary to ensure that the height of the magnetic body 1 and the upper tube 6 is the same. In this way, the magnetic body 1 can contact the second plate after being inserted into the upper tube 6. The magnetic beads can be attracted to the lower tube cover 3 by magnetic force. If more upper tubes 6 need to be designed, 2 to 3 rows of upper tubes 6 can be designed on the second plate according to the above structure. The specific number can be designed according to the actual size.

[0029] To ensure a more secure installation of the magnetic body 1, this invention also includes an upper tube cap 2 fixed to the downward-facing side of the other end of the fourth plate. The upper tube cap 2 is also cylindrical, with a height equal to or less than that of the upper tube body 6. In this case, the magnetic body 1 must be a cylinder with an outer diameter equal to the inner diameter of the upper tube body 6, thus allowing the magnetic body 1 to be securely inserted into the upper tube body 6. During a set reaction time, the magnetic bead is released by opening the upper tube cap 2. To save materials and effort, the height of the upper tube cap 2 is designed to be less than that of the upper tube body 6, so the lower end of the magnetic body 1 protrudes and is located below the upper tube cap 2.

[0030] To facilitate the fabrication of the aforementioned combined magnetic bead reaction tube, the upper tube cap 2, lower tube cap 3, lower tube body 4, first connector 5, upper tube body 6, and second connector 7 are all made of PC plastic, i.e., polycarbonate. While there are many ways to achieve the opening and closing of the first and second plates, as well as the third and fourth plates, such as the most common hinge, the aforementioned material selection makes it more convenient to integrate the first and second plates, and the third and fourth plates, into a single structure. Therefore, the aforementioned integrated structure was chosen.

[0031] The height of the upper tube 6 can be 3-8mm, while the lower tube 4, from top to bottom, includes an integral cylindrical section, a transition section, and a frustum section. Therefore, the outer diameter of the upper end of the transition section is the same as the outer diameter of the lower end of the cylindrical section, and the outer diameter of the lower end of the transition section is the same as the outer diameter of the upper end of the frustum section. The lower end of the lower tube 4 is hemispherical or conical, so the diameter of the bottom surface of the hemispherical or conical needs to be equal to the diameter of the bottom surface of the frustum section. Therefore, the inner diameter of the cylindrical section is the same as the outer diameter of the lower tube cover 3 mentioned earlier.

[0032] The magnetic material 1 may be made of, but is not limited to, ferrite magnets, rare earth magnets, AlNiCo alloy magnets and polymer magnetic compounds.

[0033] This invention relates to a combined magnetic bead reaction tube, and its use is described in different scenarios:

[0034] When performing nucleic acid extraction reactions, silanol magnetic beads are used; when performing mRNA extraction and purification, magnetic beads coated with oligonucleotides (dT) are required; when performing RPA-CRISPR reactions, magnetic beads modified with specific enzymes are required, including: first, magnetic beads for RPA reaction systems: magnetic beads modified with recombinase, magnetic beads modified with single-stranded DNA binding protein (SSB), and magnetic beads modified with strand displacement DNA polymerase; second, magnetic beads for CRISPR reaction systems: magnetic beads modified with CRISPR-CAS12 and magnetic beads modified with probes. These reactions require the addition of magnetic beads at set times and in a set order, and the release of magnetic beads according to the order required by the reaction. When the magnetic beads are not needed in the reaction, they are adsorbed back into their original positions for storage to avoid the escape of magnetic beads from the system and affect the system reaction or detection, thereby improving the reaction efficiency. Since the relevant content is all prior art, this invention will not describe it in detail. This invention uses the RPA-CRISPR reaction as an example to design a combined magnetic bead reaction tube. For other reactions, or those not mentioned, the corresponding number of upper tube caps 2 can be designed according to the same idea.

[0035] This invention discloses a combined magnetic bead reaction tube. Taking the RPA-CRISPR reaction as an example, the operation process is as follows:

[0036] Before starting the reaction, add the target bacterial strain sample to the lower tube 4, followed by 25 μL of reaction gain agent (10X) and 2 μL each of upstream and downstream primers (10 μM). Heat the lower tube 4 to 90°C and maintain this temperature for 5 minutes to facilitate cell lysis of the target bacterial strain. Start the reaction at this point. When the temperature of the lower tube 4 drops to 30°C, add the RPA reaction system magnetic beads, including magnetic beads with surface-modified recombinase, magnetic beads with surface-modified single-stranded DNA binding protein (SSB), and magnetic beads with surface-modified strand displacement DNA polymerase. Simply open one of the upper tube caps 2 above the magnetic beads to complete the addition.

[0037] After the RPA reaction is complete, another CRISPR reaction system is added, including magnetic beads with CRISPR-CAS12 surface modification and magnetic beads with probe surface modification. At this time, the other upper tube cap 2 above the magnetic beads can be opened to complete the addition. Then, the upper tube cap 2 is inserted into the upper tube body 6. When CRISPR is complete, it is necessary to adsorb the magnetic beads. Simply invert the combined magnetic bead reaction tube, and the magnetic body 1 will adsorb all the magnetic beads to complete the reaction. This achieves the function of separately adding different magnetic beads and recovering the magnetic beads.

Claims

1. A combined magnetic bead reaction tube, characterized in that, The lower tube body (4), the upper tube body (6), the first connecting piece (5) and the second connecting piece (7); The lower tube body (4) is provided with an opening, the first connecting piece (5) comprises a first plate body and a second plate body, the first plate body is sleeved above the outer wall of the lower tube body (4), the first plate body and the opening of the lower tube body (4) are flush, the first plate body extends to one side of the lower tube body (4) in the horizontal direction, the second plate body is a planar structure, the outer side of the first plate body extension section and one end of the second plate body are hingedly connected, and the outermost upper end of the first plate body extension section and the lower side of the one end of the second plate body are in contact, the second plate body is provided with a lower tube cover (3) of a cylindrical type at a position corresponding to the opening of the lower tube body (4), the outer diameter of the lower tube cover (3) is equal to the inner diameter of the opening of the lower tube body (4), and the other end of the second plate body is located at one side of the lower tube body (4). The upper tube body (6) is fixed to the second plate body at a position corresponding to the lower tube cover (3), the second connecting piece (7) comprises a third plate body and a fourth plate body, the lower end of the third plate body is fixedly connected to the other end of the second plate body, the upper end of the third plate body is away from the upper tube body (6) and flush with the upper tube body (6), one end of the fourth plate body is hingedly connected to the upper end of the third plate body, and the other end of the fourth plate body is fixedly connected to a magnetic body (1) capable of being inserted into the upper tube body (6) on one side thereof facing downward, and the magnetic body (1) has the same height as the upper tube body (6).

2. The combined magnetic bead reaction tube according to claim 1, characterized in that, The fourth plate body is further provided with a cylindrical upper tube cover (2) fixed to the other end thereof facing downward, the height of the upper tube cover (2) is equal to or less than that of the upper tube body (6), the magnetic body (1) is in the shape of a cylinder with an outer diameter equal to the inner diameter of the upper tube body (6), and the magnetic body (1) is fixedly inserted into the upper tube body (6).

3. The combined magnetic bead reaction tube according to claim 2, characterized in that, The height of the upper tube cover (2) is less than that of the upper tube body (6), and the lower end of the magnetic body (1) is located below the upper tube cover (2).

4. The combined magnetic bead reaction tube according to claim 2, wherein, The materials of the upper tube cover (2), the lower tube cover (3), the lower tube body (4), the upper tube body (6), the first connecting piece (5) and the second connecting piece (7) are all PC plastic.

5. The combined magnetic bead reaction tube according to claim 1, wherein, The magnetic body (1) is a ferrite magnet, a rare earth magnet, an aluminum-nickel-cobalt alloy magnet or a high polymer magnetic compound.

6. The combined magnetic bead reaction tube according to claim 1, wherein, The height of the upper tube body (6) is 3-8 mm.

7. The combined magnetic bead reaction tube according to claim 1, wherein, The number of the upper tube body (6) and the magnetic body (1) is the same and is several, and the several upper tube bodies (6) are uniformly arranged on the second plate body.

8. The combined magnetic bead reaction tube according to claim 1, wherein, The first plate body and the second plate body are in an integrated structure, and the third plate body and the fourth plate body are in an integrated structure.

9. The combined magnetic bead reaction tube according to claim 1, wherein, The lower tube body (4) comprises a cylindrical section, a transition section and a circular table section in an integrated structure from top to bottom, the outer diameter of the upper end of the transition section is the same as that of the lower end of the cylindrical section, the outer diameter of the lower end of the transition section is the same as that of the upper end of the circular table section, the lower end of the lower tube body (4) is in the shape of a half sphere or a circular cone, the diameter of the bottom surface of the half sphere or the circular cone is equal to that of the lower bottom surface of the circular table section, and the inner diameter of the cylindrical section is the same as the outer diameter of the lower tube cover (3).

10. A method of using a combined magnetic bead reaction tube, characterized by, A combined magnetic bead reaction tube according to any one of claims 1-9, comprising the following steps: The fourth plate body is pressed down, the magnetic body (1) is inserted into the upper tube body (6), corresponding magnetic beads are put into the lower tube cover (3) according to the reaction requirement, the magnetic body (1) adsorbs the magnetic beads through magnetic force, and then corresponding reaction is carried out; When the magnetic beads need to be added, the fourth plate body is opened, the magnetic beads enter the lower tube body (4) to participate in the reaction, after the reaction is completed, the magnetic body (1) is inserted into the upper tube body (6), the reaction tube is inverted, and all the magnetic beads are adsorbed in the lower tube cover (3).

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