Cover-opening-free reaction tube for molecular biology experiment

Abstract

The utility model discloses an uncovering-free reaction tube for molecular biology experiments, and relates to the field of biomedical treatment, the upper half part of a reaction tube body of the reaction tube is cylindrical, the upper end of the lower half part of the reaction tube is in the shape of a circular truncated cone, the diameter of the upper bottom surface of the circular truncated cone is larger than that of the lower bottom surface, and the contact parts of the upper half part and the lower half part of the reaction tube body are smoothly connected; the lining pipe comprises a cylindrical section and a circular truncated cone section, the diameter of the upper bottom surface of the circular truncated cone section is larger than that of the lower bottom surface, the lower end of the cylindrical section is smoothly connected with the upper end of the circular truncated cone section, and the outer wall of the cylindrical section of the lining pipe is attached to the inner wall of the upper half part of the reaction pipe body; the outer wall of the circular truncated cone section of the lining pipe is attached to the inner wall of the upper end part in the lower half part of the reaction pipe body; a liquid adding through hole and a plurality of liquid storage compartments with the same depth are formed in the lining pipe in the vertical direction, the upper ends of the liquid storage compartments are open, the bottom face of each liquid storage compartment extends downwards in the vertical direction to be provided with a corresponding through hole, and the through holes are filled with paraffin.

Description

Technical Field

[0001] The utility model relates to the field of biological medicine, in particular to a reaction tube for molecular biology experiments which does not require opening of a cover. Background Art

[0002] Microcentrifuge tubes are a type of experimental consumables that are frequently used in reactions such as nucleic acid extraction, immunology, and molecular purification. Since their invention in the 1960s, they have been widely used in immunoassays, immunodetection, cell separation, biomacromolecule purification, and molecular biology.

[0003] With the continuous development of molecular biology experimental techniques and methods, the number of methods that require opening the lid in the middle of the experimental process to add reagents is gradually increasing. Taking nucleic acid detection experiments as an example, in the RPA-CRISPR-CAS detection method, the RPA system that has completed the first step of the reaction needs to be opened in the middle of the process and added to the CRISPR-CAS system for the next step of the reaction. However, this type of operation requires frequent opening of the lid during the reaction, which will not only contaminate the samples in the second step of the reaction, but may also form aerosol pollution, affecting the subsequent reaction experiments. For nucleic acid detection experiments with biohazardous samples, such as virus detection, the released aerosols will cause great harm to the personal health and safety of the operators. At present, when solving these problems, technicians often need to use specially customized reaction equipment or reagents, but such solutions are costly and complex in structure. Utility Model Content

[0004] In view of the problems existing in the prior art, the utility model provides a reaction tube for molecular biology experiments that does not require opening of the cover, which can release multiple reagents in batches without opening the cover, will not contaminate the reaction samples, will not form aerosol pollution, and ensures the smooth progress of subsequent reaction experiments.

[0005] The utility model is realized by the following technical solutions:

[0006] A reaction tube for molecular biology experiments that does not require opening of the cover, comprising a reaction tube and an inner liner tube;

[0007] The upper half of the reaction tube body of the reaction tube is cylindrical, the upper end of the lower half of the reaction tube body is truncated, the upper bottom surface diameter of the truncated cone is larger than the lower bottom surface diameter, and the contact portion between the upper half and the lower half of the reaction tube body is smoothly connected;

[0008] The inner liner tube comprises a cylindrical section and a truncated cone section, the upper bottom surface diameter of the truncated cone section is larger than the lower bottom surface diameter, the lower end of the cylindrical section is smoothly connected to the upper end of the truncated cone section, the outer wall of the cylindrical section of the inner liner tube is attached to the inner wall of the upper half of the reaction tube body, and the outer wall of the truncated cone section of the inner liner tube is attached to the inner wall of the upper end portion of the lower half of the reaction tube body;

[0009] The inner liner tube is provided with a liquid adding through hole and a plurality of liquid storage compartments of the same depth in the vertical direction. The upper end of the liquid storage compartment is open. The bottom surface of each liquid storage compartment is provided with a corresponding through hole extending downward in the vertical direction. The through hole is filled with paraffin.

[0010] Preferably, the reaction tube further comprises a tube cover, the reaction tube body comprises a cylindrical section and a truncated cone section of an integral structure from top to bottom, the lower end of the reaction tube body is semi-spherical or conical, and the diameter of the bottom surface of the semi-spherical or conical is equal to the diameter of the lower bottom surface of the truncated cone section;

[0011] One end of the tube cover is movably connected to one side of the upper end surface of the cylindrical section, and the lower end surface of the other end of the tube cover extends downward in the vertical direction to be provided with a convex ring with an outer diameter equal to the inner diameter of the cylindrical section, and the convex ring is inserted at the opening of the cylindrical section.

[0012] Furthermore, in the longitudinal section of the reaction tube body along its own central axis, the angle between the straight lines corresponding to the cylindrical segment and the truncated cone segment located on the same side is 150 to 170 degrees.

[0013] Furthermore, the height dimension of the truncated cone section of the inner liner tube is twice the height dimension of the lower half of the reaction tube body.

[0014] Furthermore, the height dimension of the cylindrical section of the inner liner tube is

[0015] Preferably, the number of the liquid storage compartments is 1 to 6.

[0016] Furthermore, the cross sections of the liquid adding through hole and all the liquid storage compartments are all fan-shaped with equal curvature.

[0017] Furthermore, the liquid adding through holes and all the liquid storage compartments are evenly distributed along the cross section of the inner liner tube.

[0018] Furthermore, a cylindrical through hole is provided at the center of each liquid storage compartment.

[0019] Preferably, after the through holes are filled with paraffin, a paraffin layer is formed on the bottom surface of each liquid storage compartment, the paraffin layer is in contact with the inner wall of the liquid storage compartment, and the thickness of the paraffin layer is twice the depth of the liquid storage compartment.

[0020] Compared with the prior art, the utility model has the following beneficial technical effects:

[0021] The utility model discloses a reaction tube for molecular biology experiments that does not require opening of the cover. The upper bottom surface diameter of the truncated cone-shaped part at the upper end of the lower half of the reaction tube body is larger than the lower bottom surface. This design facilitates the clamping of an inner liner tube of the same shape at the corresponding position of its inner wall to maintain vertical stability when it is smoothly connected at the contact point with the upper half of the reaction tube body. The reaction reagent can be added in a suitable time period according to the reaction needs through the inner liner tube. Paraffin is a heat-sensitive material that has the property of undergoing phase change with temperature changes (such as melting or solidification). When an external heat source heats the reaction tube, the paraffin will gradually melt, and the reaction system added thereto will flow into the main reaction system along the through hole, thereby participating in the reaction. The vertical liquid adding through hole can add the reactants or reagents required for the main reaction system to the lower half of the reaction tube body in advance. The reaction tube of the utility model realizes the isolation and mixing control between different reactants, and can release multiple reagents in batches without opening the cover, so that the reaction process can be carried out in stages, which is helpful to improve the reaction efficiency, product purity and process safety, and will not cause the reaction sample to be contaminated, and will not form aerosol pollution, thereby ensuring the smooth progress of subsequent reaction experiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is a schematic diagram of the external structure of the reaction tube of the utility model.

[0023] Figure 2 It is a schematic diagram of the internal section of the reaction tube of the utility model.

[0024] Figure 3 yes Figure 1 Schematic diagram of the inner liner pipe.

[0025] Figure 4 yes Figure 1 Top view of the inner liner pipe.

[0026] Figure 5 yes Figure 1 An angled cross-sectional diagram of the inner liner pipe.

[0027] Figure 6 yes Figure 1 Another angle cross-sectional diagram of the inner liner pipe.

[0028] Figure 7 yes Figure 6 Schematic diagram of the cross section of the middle liner pipe after sealing treatment.

[0029] Figure 8 yes Figure 5 Schematic diagram of the cross section of the middle liner pipe after sealing treatment.

[0030] In the figure: 101 - tube cover, 102 - reaction tube body, 103 - liner tube, 1031 - liquid adding through hole, 1032 - liquid storage compartment, 1033 - through hole, 1034 - sealing layer. DETAILED DESCRIPTION

[0031] The present invention is further described below in conjunction with the accompanying drawings and embodiments, but they are not intended to limit the present invention.

[0032] The utility model discloses a reaction tube for molecular biology experiments without opening the cover. Figure 1 and Figure 2 As shown, it includes a tube cover 101, a reaction tube body 102 and an inner liner 103. The reaction tube body 102 includes a cylindrical section (upper half) and a truncated cone section (lower half) of an integral structure from top to bottom, which is used to carry the reaction system. It is designed using a common structure. The upper bottom surface diameter of the truncated cone is larger than the lower bottom surface diameter, and the contact between the cylindrical section and the truncated cone section is smoothly connected. One end of the tube cover 101 is movably connected to one side of the upper end surface of the cylindrical section, and the lower end surface of the other end extends downward in the vertical direction and is provided with a convex ring with an outer diameter equal to the inner diameter of the cylindrical section, and the convex ring is plugged into the opening of the cylindrical section. The lower end of the reaction tube body 102 is semi-spherical or conical, and the diameter of the semi-spherical or conical bottom surface is equal to the diameter of the lower bottom surface of the truncated cone section.

[0033] like Figure 2 and Figure 3 As shown, the inner liner tube 103 is composed of a cylindrical section and a truncated cone section. The diameter of the upper bottom surface of the truncated cone section is larger than the diameter of the lower bottom surface. The lower end of the cylindrical section is smoothly connected to the upper end of the truncated cone section. The outer wall of the cylindrical section of the inner liner tube 103 is attached to the inner wall of the cylindrical section of the reaction tube body 102, and the outer wall of the truncated cone section is attached to the inner wall of the upper end of the truncated cone section of the reaction tube body 102.

[0034] like Figure 4 As shown, the inner liner 103 has a liquid adding hole 1031 and 1 to 6 liquid storage compartments 1032 with the same depth in the vertical direction. The cross-sections of the liquid adding hole 1031 and all the liquid storage compartments 1032 are all sectors with equal arcs. The upper ends of the liquid storage compartments 1032 are open. The liquid adding hole 1031 and all the liquid storage compartments 1032 are evenly distributed along the cross-section of the inner liner 103. Figure 5 and Figure 6 As shown, a corresponding cylindrical through hole 1033 is provided at the center of the bottom surface of each liquid storage compartment 1032 extending downward in the vertical direction. After each through hole 1033 is filled with different types of paraffin wax, as shown in FIG. Figure 7 and Figure 8 As shown, a paraffin layer is formed on the bottom surface of each liquid storage compartment 1032, and the paraffin layer is in contact with the inner wall of the liquid storage compartment 1032. The thickness of the paraffin layer is equal to the depth of the liquid storage compartment 1032. The paraffin layer and the paraffin in the through hole 1033 form a sealing layer 1034, which can seal the through hole 1033 and separate the reaction liquid in the liquid storage compartment 1032 from the main system.

[0035] Specifically, in the longitudinal section of the reaction tube body 102 along its own central axis, the angle between the straight lines corresponding to the cylindrical section and the truncated cone section on the same side is 150-170 degrees, and the height of the truncated cone section of the liner tube 103 is 1 / 2 of the height of the truncated cone section of the reaction tube body 102. The height of the cylindrical section of the liner tube 103 is the height of the cylindrical section of the reaction tube body 102.

[0036] Specifically, when closing the through hole 1033, first, place the conical section of the liner tube 103 on a clean operating platform, and add pre-melted paraffin oil of different brands into the liquid storage compartment 1032 of the liner tube 103 as needed, ensuring that the paraffin oil covers the through hole 1033 and reaches the expected height on the inner wall of the liquid storage compartment 1032, wait for the paraffin oil to solidify, and install the closed liner tube 103 into the reaction tube body 102.

[0037] When there is one liquid storage compartment 1032, RPA-CRISPR-CAS reaction can be performed, and the corresponding paraffin that can be used is 40# paraffin.

[0038] When there are two liquid storage compartments 1032, RPA-Ago reaction can be performed, and the paraffins that can be used are 30# paraffin and 80# paraffin.

[0039] When there are three liquid storage compartments 1032, RT-RPA-Ago reaction can be performed, and the paraffins that can be used are 60# paraffin and 80# paraffin.

[0040] When there are four liquid storage compartments 1032, RT-RPA-LAMP-Ago reaction can be performed, and the corresponding paraffins that can be used are 30# paraffin, 40# paraffin, 60# paraffin, and 80# paraffin.

[0041] The utility model discloses a reaction tube for molecular biology experiment without opening the cover. When carrying out a specific molecular biology experiment, the operation process is as follows:

[0042] First, the first step reaction system is added into the reaction tube body 102 through the liquid adding through hole 1031 , and the required reaction systems are added into the liquid storage compartment 1032 . After completion, the reaction tube body 102 is closed by the tube cover 101 .

[0043] Afterwards, when the reaction is carried out in the reaction tube 102, the process is as follows:

[0044] First, the main reaction system is reacted at the reaction temperature required by the main reaction system. At this time, the reaction temperature of the main reaction system is lower than the melting point of all the paraffins serving as the closed layer 1034. After the reaction of the main reaction system is completed, the reaction tube is heated to the melting point of the paraffin corresponding to the closed layer 1034, and then quickly centrifuged. After centrifugation, the corresponding paraffin oil floats on the upper layer, and a second reaction system is formed in the reaction tube and the second step reaction begins. If there are a third reaction system and a fourth reaction system, the process is the same as the second reaction system, and the key point is that the temperature during the reaction is lower than the melting point of all the paraffins in the remaining closed layers 1034, and then the melting point of the paraffin corresponding to the closed layer 1034 is heated until the reaction is completed.

[0045] Example 1

[0046] When performing the RPA-CRISPR-CAS reaction, there is one liquid storage compartment 1032, numbered as liquid storage compartment 1, and the lower half of the reaction tube body 102 is placed with relevant enzymes, primers, probes, reaction enhancers and water for the RPA amplification system.

[0047] A layer of 40# paraffin is filled in the through hole of the liquid storage chamber 1, and the CRISPR-CAS detection system is added to the chamber storage 1. When the amplification reaction is completed, the temperature is briefly raised to 40°C, and the through hole is opened after the paraffin melts. The tube body is centrifuged to allow the CRISPR-CAS detection system to enter the lower half of the reaction tube body 102, thereby participating in the reaction. The CRISPR-CAS detection system is mixed with the amplified RPA system, and the system starts to react at 35-42°C and releases a signal.

[0048] Example 2

[0049] When performing the RT-LAMP-pfAgo reaction, there are three liquid storage compartments 1032, numbered as liquid storage compartment 1, liquid storage compartment 2 and liquid storage compartment 3. The relevant enzymes, primers, probes, reaction enhancers and water used in the RT-LAMP amplification system are placed in the lower half of the reaction tube body 102.

[0050] A layer of 60# paraffin is filled in the through hole of the liquid storage chamber 1, where the template for nucleic acid amplification is stored. When the temperature reaches the reaction temperature of RT-LAMP 65°C, the paraffin melts and the through hole opens. The tube body is centrifuged to allow the template for nucleic acid amplification to enter the lower half of the reaction tube body 102 to participate in the reaction.

[0051] A layer of 80# paraffin was filled in the through hole between liquid storage chamber 2 and liquid storage chamber 3. pfAgo protein, reaction buffer, 5' phosphorylated gDNA, and molecular beacons for releasing signals were stored in liquid storage chamber 2. Mn 2+When the reaction temperature reaches 80°C, the paraffin in liquid storage chamber 2 and liquid storage chamber 3 melts, opening their respective through holes and then centrifuging. The pfAgo reaction system is mixed with the amplified RT-LAMP system. When the temperature reaches 95°C, the pfAgo reaction begins and the signal is released.

Claims

1. A reaction tube for molecular biology experiments without opening the cover, characterized in that: It includes a reaction tube and an inner liner tube (103); The upper half of the reaction tube body (102) of the reaction tube is cylindrical, the upper end of the lower half of the reaction tube body (102) is truncated, the upper bottom surface diameter of the truncated cone is larger than the lower bottom surface diameter, and the contact portion between the upper half and the lower half of the reaction tube body (102) is smoothly connected; The inner liner tube (103) comprises a cylindrical section and a truncated cone section, the upper bottom surface diameter of the truncated cone section is larger than the lower bottom surface diameter, the lower end of the cylindrical section is smoothly connected to the upper end of the truncated cone section, the outer wall of the cylindrical section of the inner liner tube (103) is attached to the inner wall of the upper half of the reaction tube body (102), and the outer wall of the truncated cone section of the inner liner tube (103) is attached to the inner wall of the upper end portion of the lower half of the reaction tube body (102); The inner liner tube (103) is provided with a liquid adding through hole (1031) and a plurality of liquid storage compartments (1032) of the same depth in the vertical direction. The upper end of the liquid storage compartment (1032) is open, and the bottom surface of each liquid storage compartment (1032) is provided with a corresponding through hole (1033) extending downward in the vertical direction. The through hole (1033) is filled with paraffin.

2. The cover-free reaction tube for molecular biology experiments according to claim 1, characterized in that: The reaction tube further comprises a tube cover (101); the reaction tube body (102) comprises a cylindrical section and a truncated cone section of an integral structure from top to bottom; the lower end of the reaction tube body (102) is semi-spherical or conical, and the diameter of the bottom surface of the semi-spherical or conical section is equal to the diameter of the lower bottom surface of the truncated cone section; One end of the tube cover (101) is movably connected to one side of the upper end surface of the cylindrical section, and the lower end surface of the other end of the tube cover (101) extends downward in the vertical direction and is provided with a convex ring with an outer diameter equal to the inner diameter of the cylindrical section, and the convex ring is inserted into the opening of the cylindrical section.

3. The cover-free reaction tube for molecular biology experiments according to claim 2, characterized in that: In the longitudinal section of the reaction tube body (102) along its own central axis, the angle between the straight lines corresponding to the cylindrical segment and the truncated cone segment located on the same side is 150 to 170 degrees.

4. The cover-free reaction tube for molecular biology experiments according to claim 2, characterized in that: The height dimension of the truncated cone section of the inner liner tube (103) is equal to the height dimension of the lower half of the reaction tube body (102).

5. The cover-free reaction tube for molecular biology experiments according to claim 2, characterized in that: The height dimension of the cylindrical section of the inner lining tube (103) is equal to the height dimension of the cylindrical section of the reaction tube body (102).

6. The cover-free reaction tube for molecular biology experiments according to claim 1, characterized in that: The number of the liquid storage compartments (1032) is 1 to 6.

7. The cover-free reaction tube for molecular biology experiments according to claim 6, characterized in that: The cross sections of the liquid adding through hole (1031) and all the liquid storage compartments (1032) are all fan-shaped with equal curvature.

8. The cover-free reaction tube for molecular biology experiments according to claim 7, characterized in that: The liquid adding through holes (1031) and all the liquid storage compartments (1032) are evenly distributed along the cross section of the inner lining tube (103).

9. The cover-free reaction tube for molecular biology experiments according to claim 7, characterized in that: A cylindrical through hole (1033) is provided at the center of each liquid storage compartment (1032).

10. The cover-free reaction tube for molecular biology experiments according to claim 1, characterized in that: After the through hole (1033) is filled with paraffin, a paraffin layer is formed on the bottom surface of each liquid storage compartment (1032), and the paraffin layer is in contact with the inner wall of the liquid storage compartment (1032). The thickness of the paraffin layer is equal to the depth of the liquid storage compartment (1032).

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