A gene knockout plasmid kit

Abstract

The utility model discloses a gene knockout plasmid kit applied to the field of reagent kit, including outer protective pipe, detachable embedding in the inner tube of outer protective pipe and cover outer protective pipe, the sealing cover of inner tube opening, the inner side fixed limit hoop ring of outer protective pipe, and the bottom extends to the bottom of outer protective pipe, and the intermediate layer is formed between outer protective pipe and inner tube, the upper end opening of outer protective pipe integrative has the connecting ring, and the inner thread portion is provided with in connecting ring inner wall, and the outer thread structure of thread adaptation with inner thread portion is formed to sealing cover circumferential side, through the sealing cover is screwed in connecting ring, makes the partition ring insert between two elastic self -sealing ring, and partition ring lower extreme and limit hoop ring abut tightly, like this can realize multiple sealing effect, and the overall sealing effect improves greatly, is more favorable to the storage security of gene knockout plasmid is improved, and can provide more obvious prompt effect through the prompt component of setting.

Description

Technical Field

[0001] This invention relates to the field of reagent kits, and in particular to a gene knockout plasmid reagent kit. Background Technology

[0002] pLenti-METTL3-sgRNA (METTL3 gene knockout plasmid) is a plasmid that can simultaneously express Cas9, the target gene's sgRNA, and the puromycin resistance gene in animal cells. It is used to directly knock out the target gene in animal cells using CRISPR / Cas9 technology, or to knock out the target gene using CRISPR / Cas9 technology after packaging lentiviruses.

[0003] In the prior art, a special kit (double-layered tubular reagent tube) is usually used to store the gene knockout plasmid, which is similar to a kit for knocking out the DHC2 gene in glioblastoma cells disclosed in the patent document with publication number "CN107841509B".

[0004] In actual use, it was found that although the existing kits can meet the basic storage requirements, the double-layer tubes do not form a good isolation seal. Therefore, when the outer tube is not sealed tightly, the inner tube is also affected, which is not conducive to the safe storage of gene knockout plasmids. At the same time, there is no corresponding indication when the outer tube is not sealed well, which makes it difficult for users to judge the storage status. Utility Model Content

[0005] The core of this invention lies in solving the problem of inadequate isolation and sealing between double-layered pipes in existing technologies by using a combination of an elastic self-sealing ring, a partition ring, a limiting ring, and a sealing plug. Simultaneously, the included indicator component reacts with air and changes color when air enters between the inner side of the partition ring and the outer side of the inner pipe, providing a clear indication effect.

[0006] To solve the above problems, the present invention adopts the following technical solution.

[0007] A gene knockout plasmid kit includes an outer sheath, a removable inner tube embedded in the outer sheath, and a sealing cap that covers the openings of the outer sheath and the inner tube.

[0008] A limiting ring is fixed inside the outer protective tube, and the inner tube is inserted into the limiting ring with its bottom extending to the bottom of the outer protective tube, forming a middle partition layer between the outer protective tube and the inner tube.

[0009] The upper opening of the outer protective tube is integrally formed with a connecting ring, the inner wall of the connecting ring is provided with an internal thread, and the periphery of the sealing cap is formed with an external thread structure that matches the thread of the internal thread.

[0010] Both the outer side of the upper end of the inner tube and the inner wall of the connecting ring are fixed with elastic self-sealing rings, and the two elastic self-sealing rings move and press against each other on the side closest to each other.

[0011] The lower end of the sealing cap has a convex ring that is embedded in the gap between the upper end of the inner tube and the connecting ring. The bottom of the convex ring of the sealing cap is rotatably connected to a partition ring. The lower end of the partition ring moves through the space between two elastic self-sealing rings and abuts against the top of the limiting ring. The lower end of the sealing cap is also fixed with a sealing plug. The sealing plug is placed in the upper opening of the inner tube and forms a seal.

[0012] An indicator component is installed on the outer side of the inner tube above the limiting ring. The indicator component reacts with air and changes color when air enters between the inner side of the isolation ring and the outer side of the inner tube.

[0013] By tightening the sealing cap inside the connecting ring, the partition ring is inserted between the two elastic self-sealing rings, and its lower end abuts against the limiting ring. This creates a first seal between the lower end of the sealing cap and the outer protective tube, a second seal between the outer elastic self-sealing ring and the partition ring, a third seal between the partition ring and the limiting ring, and a fourth seal between the inner elastic self-sealing ring and the partition ring. Simultaneously, the sealing plug is inserted into the opening of the inner tube to create a fifth seal. This achieves multiple sealing effects, significantly improving the overall sealing performance and enhancing the storage safety of gene knockout plasmids.

[0014] In addition, the indicator component can react with air and change color when air enters between the inside of the partition ring and the outside of the inner tube. Users can then visually observe whether there is any lack of airtightness through the transparent outer protective tube and partition ring, which allows for a more intuitive assessment of the storage status of the gene knockout plasmid, resulting in better actual use.

[0015] Furthermore, a pipe cap sleeve is fitted on the outside of the connecting ring, and a rotating connecting ring is rotatably fitted on the upper end of the sealing cap. The sealing cap is rotatably connected to the pipe cap sleeve through the rotating connecting ring, and the lower end of the rotating connecting ring abuts against the top of the connecting ring.

[0016] Furthermore, a limiting protrusion is integrally formed on the outer side of the inner tube, with the bottom of the limiting protrusion abutting against the top of the limiting hoop, and the outer diameter of the limiting protrusion being smaller than the inner diameter of the partition ring.

[0017] Furthermore, the prompting component includes a fixing box, a color-changing body, and a sliding cover. The fixing box is fixed to the outside of the inner tube, the color-changing body is fixed inside the fixing box, the sliding cover is slidably connected to the fixing box and closes the opening of the fixing box, and a compression spring is fixed to the lower end of the fixing box. The lower end of the compression spring is fixed to the top of the limiting protrusion ring.

[0018] Furthermore, the partition ring has an integrally formed driving rib on the inner side near the inner tube. The driving rib is a strip-shaped protrusion that protrudes towards one side of the inner tube. The sliding cover is an L-shaped ring with the top of the ring at the lower end of the sliding cover abutting against the bottom of the driving rib.

[0019] Furthermore, a sealing gasket is fixed to the top of the sliding cover. When the sliding cover closes the fixing box, the top of the sealing gasket abuts against the top wall inside the fixing box.

[0020] Compared with existing technologies, the advantages of this utility model are:

[0021] This solution achieves multiple sealing effects by tightening the sealing cap inside the connecting ring, allowing the partition ring to insert between the two elastic self-sealing rings, with the lower end of the partition ring abutting against the limiting ring. This creates a first seal between the lower end of the sealing cap and the outer protective tube, a second seal between the elastic self-sealing ring and the partition ring, a third seal between the partition ring and the limiting ring, and a fourth seal between the elastic self-sealing ring and the partition ring. Simultaneously, the sealing plug is inserted into the opening of the inner tube to create a fifth seal. This achieves multiple sealing effects, significantly improving the overall sealing performance and enhancing the storage safety of gene knockout plasmids.

[0022] This solution features a notification component that reacts with air and changes color when air enters between the inner side of the partition ring and the outer side of the inner tube. Users can then visually observe whether the seal is not tight through the transparent outer protective tube and partition ring, allowing for a more intuitive assessment of the storage status of the gene knockout plasmid and resulting in better practical performance. Attached Figure Description

[0023] Figure 1 This is a three-dimensional structural diagram of the present invention;

[0024] Figure 2 This is a schematic diagram of the outer protective tube and inner tube structure of this utility model;

[0025] Figure 3 This is a schematic diagram of the overhead side structure of the sealing cap and pipe cover sleeve of this utility model;

[0026] Figure 4 This is a cross-sectional view of the present invention;

[0027] Figure 5 for Figure 4 Enlarged view of the structure at point A in the middle.

[0028] Explanation of the labels in the diagram:

[0029] 1. Outer protective tube; 11. Connecting ring; 111. Internal threaded part; 12. Limiting hoop ring; 2. Inner tube; 21. Limiting convex ring; 3. Sealing cap; 31. Partition ring; 311. Driving rib; 32. Sealing plug; 4. Pipe cap sleeve; 5. Rotating connecting ring; 6. Elastic self-sealing ring; 7. Indicator component; 71. Fixing box; 72. Color-changing body; 73. Sliding cover; 74. Compression spring; 8. Sealing gasket; 9. Anti-slip ring. Detailed Implementation

[0030] The technical solution will now be clearly and completely described with reference to the accompanying drawings in the embodiments of this utility model.

[0031] First implementation method:

[0032] Please see Figures 1-3 A gene knockout plasmid kit includes an outer sheath 1, an inner tube 2 that is removably embedded in the outer sheath 1, and a sealing cap 3 that covers the openings of the outer sheath 1 and the inner tube 2.

[0033] A limiting ring 12 is fixed inside the outer protective tube 1, and the inner tube 2 is inserted into the limiting ring 12 and extends to the bottom of the outer protective tube 1. A middle partition layer is formed between the outer protective tube 1 and the inner tube 2.

[0034] The upper opening of the outer protective tube 1 is integrally formed with a connecting ring 11, the inner wall of the connecting ring 11 is provided with an internal thread 111, and the periphery of the sealing cover 3 is formed with an external thread structure that is adapted to the thread of the internal thread 111.

[0035] Elastic self-sealing rings 6 are fixed on the outer side of the upper end of the inner tube 2 and on the inner wall of the connecting ring 11. The two elastic self-sealing rings 6 move and press against each other on the side that is close to each other.

[0036] The lower end of the sealing cap 3 has a convex ring that is embedded in the gap between the upper end of the inner tube 2 and the connecting ring 11. The bottom of the convex ring of the sealing cap 3 is rotatably connected to the partition ring 31. The lower end of the partition ring 31 moves through the gap between the two elastic self-sealing rings 6 and abuts against the top of the limiting ring 12. The lower end of the sealing cap 3 is also fixed with a sealing plug 32. The sealing plug 32 is inserted into the upper opening of the inner tube 2 and forms a seal.

[0037] Based on the above structure, after the sealing cap 3 is tightened inside the connecting ring 11, the partition ring 31 is inserted between the two elastic self-sealing rings 6, and the lower end of the partition ring 31 abuts against the limiting ring 12, so that the lower end of the sealing cap 3 and the outer protective tube 1 form a first sealing point, the outer elastic self-sealing ring 6 (fixed with the outer protective tube 1) and the partition ring 31 form a second sealing point, the partition ring 31 and the limiting ring 12 form a third sealing point, and the inner elastic self-sealing ring 6 (fixed with the inner tube 2) and the partition ring 31 form a fourth sealing point. At the same time, the sealing plug 32 is inserted into the opening of the inner tube 2 to form a fifth sealing point. In this way, multiple sealing effects can be achieved, the overall sealing effect is greatly improved, and it is more conducive to improving the storage safety of gene knockout plasmids.

[0038] Furthermore, a pipe cap sleeve 4 is fitted on the outside of the connecting ring 11, and a rotating connecting ring 5 is rotatably fitted on the upper end of the sealing cap 3. The sealing cap 3 is rotatably connected to the pipe cap sleeve 4 through the rotating connecting ring 5, and the lower end of the rotating connecting ring 5 is pressed against the top of the connecting ring 11.

[0039] The tube cap sleeve 4 further protects the connecting ring 11 and forms a new outermost sealing layer, further improving the sealing effect of the gene knockout plasmid kit.

[0040] Furthermore, an integrally formed limiting protrusion ring 21 is formed on the outer side of the inner tube 2. The bottom of the limiting protrusion ring 21 abuts against the top of the limiting hoop ring 12. The outer diameter of the limiting protrusion ring 21 is smaller than the inner diameter of the partition ring 31, which facilitates the positioning of the inner tube 2 when it is placed into the outer protective tube 1, preventing the inner tube 2 from being unable to "suspend" in the outer protective tube 1. This creates an effective separation and buffer space between the outer protective tube 1 and the inner tube 2, providing good protection for the inner tube 2.

[0041] In addition, in order to facilitate gripping of the outer protective tube 1, an anti-slip ring 9 is fixedly fitted on the outside of the outer protective tube 1, and the outer surface of the anti-slip ring 9 is provided with anti-slip texture.

[0042] Second implementation method:

[0043] Please see Figures 3-5 A gene knockout plasmid kit, which differs from Example 1 in that:

[0044] An indicator component 7 is installed on the outer side of the inner tube 2 above the limiting ring 12. The indicator component 7 reacts with air and changes color when air enters between the inner side of the isolation ring 31 and the outer side of the inner tube 2.

[0045] The indicator component 7 is designed to react with air and change color when air enters between the inner side of the isolation ring 31 and the outer side of the inner tube 2. Users can then visually observe whether there is any lack of airtightness through the transparent outer protective tube 1 and the isolation ring 31, which allows for a more intuitive assessment of the storage status of the gene knockout plasmid and results in better actual use.

[0046] Specifically, the prompt component 7 includes a fixed box 71, a color-changing body 72, and a sliding cover 73. The fixed box 71 is fixed to the outside of the inner tube 2. The color-changing body 72 is fixed inside the fixed box 71. The color-changing body 72 is a transparent porous capsule containing a color-changing substance. The color-changing substance refers to a substance commonly used in the prior art and applicable to this embodiment that can react with oxygen in the air, such as copper powder (which oxidizes to basic copper carbonate, which is green) or phenolic compounds (such as phenol, which, when oxidized in the air, generates quinone compounds, causing the color to change from colorless to pink or dark). The sliding cover 73 is slidably connected to the fixed box 71 and closes the opening of the fixed box 71. A compression spring 74 is fixed to the lower end of the fixed box 71. The lower end of the compression spring 74 is fixed to the top of the limiting protrusion ring 21. Both the fixed box 71 and the sliding cover 73 are made of materials that do not react with the color-changing substance, such as polytetrafluoroethylene (PTFE) or glass. Considering the actual cost and the storage requirements of the gene knockout plasmid, polytetrafluoroethylene is preferred in this embodiment.

[0047] The partition ring 31 has an integrally formed driving rib 311 on the inner side near the inner tube 2. The driving rib 311 is a strip-shaped protrusion protruding to one side of the inner tube 2. The sliding cover 73 is an L-shaped ring body. The top of the ring body at the lower end of the sliding cover 73 abuts against the bottom of the driving rib 311.

[0048] When the partition ring 31 is inserted downward between the two elastic self-sealing rings 6, the driving rib 311 on the inner side of the partition ring 31 abuts against the lower end ring of the sliding cover 73, thereby driving the sliding cover 73 to move downward against the elastic force of the compression spring 74, thus opening the sliding cover 73 and exposing the color-changing body 72 in the cavity between the outer side of the inner tube 2 and the inner side of the partition ring 31. This allows the color-changing body 72 to react with the air and change color when the air enters the cavity between the outer side of the inner tube 2 and the inner side of the partition ring 31, thus playing a corresponding indication role.

[0049] Furthermore, a sealing gasket 8 is fixed to the top of the sliding cover 73. The sealing gasket 8 is a rubber gasket, and there is a gap between its inner side and the color-changing body 72 inside the fixed box 71 to prevent the two from reacting. When the sliding cover 73 closes the fixed box 71, the top of the sealing gasket 8 is pressed against the inner top wall of the fixed box 71. Through the setting of the sealing gasket 8, the sliding cover 73 can form a sealed box when closing the fixed box 71, ensuring that the color-changing body 72 inside will not come into contact with the outside air, so that the inner tube 2 will not be wasted when it is taken out for use alone.

[0050] The above description is merely a preferred embodiment of this utility model; it encompasses all the protection scope of this utility model. Any equivalent substitutions or modifications made by those skilled in the art within the technical scope disclosed in this utility model, based on the technical solution and its improved concept, should be covered within the protection scope of this utility model.

Claims

1. A gene knockout plasmid kit, comprising an outer sheath (1), an inner tube (2) removably embedded within the outer sheath (1), and a sealing cap (3) covering the openings of the outer sheath (1) and the inner tube (2), characterized in that: The outer protective tube (1) is fixed with a limiting ring (12) on the inner side, and the inner tube (2) is inserted into the limiting ring (12) and extends to the bottom of the outer protective tube (1). A middle partition layer is formed between the outer protective tube (1) and the inner tube (2). The upper opening of the outer protective tube (1) is integrally formed with a connecting ring (11), the inner wall of the connecting ring (11) is provided with an internal thread (111), and the periphery of the sealing cover (3) is formed with an external thread structure that is adapted to the internal thread (111). Both the outer side of the upper end of the inner tube (2) and the inner wall of the connecting ring (11) are fixed with elastic self-sealing rings (6), and the two elastic self-sealing rings (6) move and abut against each other on the side that is close to each other. The lower end of the sealing cap (3) has a convex ring that is embedded in the gap between the upper end of the inner tube (2) and the connecting ring (11). The bottom of the convex ring of the sealing cap (3) is rotatably connected to a partition ring (31). The lower end of the partition ring (31) moves through the gap between two elastic self-sealing rings (6) and abuts against the top of the limiting ring (12). The lower end of the sealing cap (3) is also fixed with a sealing plug (32). The sealing plug (32) is inserted into the upper opening of the inner tube (2) and forms a seal. An indicator component (7) is installed on the outer side of the inner tube (2) above the limiting ring (12). The indicator component (7) reacts with air and changes color when air enters between the inner side of the isolation ring (31) and the outer side of the inner tube (2).

2. The gene knockout plasmid kit according to claim 1, characterized in that: The outer side of the connecting ring (11) is fitted with a pipe cap sleeve (4), and the upper end of the sealing cap (3) is rotatably fitted with a rotating connecting ring (5). The sealing cap (3) is rotatably connected to the pipe cap sleeve (4) through the rotating connecting ring (5), and the lower end of the rotating connecting ring (5) is pressed against the top of the connecting ring (11).

3. The gene knockout plasmid kit according to claim 2, characterized in that: The inner tube (2) has an integrally formed limiting protrusion ring (21) on the outside. The bottom of the limiting protrusion ring (21) abuts against the top of the limiting hoop ring (12). The outer diameter of the limiting protrusion ring (21) is smaller than the inner diameter of the partition ring (31).

4. A gene knockout plasmid kit according to claim 3, characterized in that: The prompting component (7) includes a fixed box (71), a color-changing body (72), and a sliding cover (73). The fixed box (71) is fixed to the outside of the inner tube (2). The color-changing body (72) is fixed inside the fixed box (71). The sliding cover (73) is slidably connected to the fixed box (71) and closes the opening of the fixed box (71). A compression spring (74) is fixed at the lower end of the fixed box (71). The lower end of the compression spring (74) is fixed to the top of the limiting protrusion (21).

5. A gene knockout plasmid kit according to claim 4, characterized in that: The partition ring (31) has an integrally formed drive rib (311) on the inner side near the inner tube (2). The drive rib (311) is a strip-shaped protrusion that protrudes towards one side of the inner tube (2). The sliding cover (73) is an L-shaped ring. The top of the ring at the lower end of the sliding cover (73) abuts against the bottom of the drive rib (311).

6. A gene knockout plasmid kit according to claim 4, characterized in that: A sealing gasket (8) is fixed to the top of the sliding cover (73). When the sliding cover (73) closes the fixing box (71), the top of the sealing gasket (8) abuts against the inner top wall of the fixing box (71).

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