Efficient nucleic acid extraction and adsorption column

Abstract

The invention relates to the technical field of nucleic acid extraction, in particular to an efficient nucleic acid extraction and adsorption column which comprises a treatment cylinder, a mixed cracking unit, an extraction cylinder, a release unit and an elution cylinder, the problems that in the prior art, when nucleic acid is extracted, a transfer pipette cannot be sealed, so that a sample and a lysis solution are prone to leakage and pollution in the mixing process, and the purity of the nucleic acid is affected can be solved; the adsorption column needs to be transferred into other containers for subsequent washing purification and elution collection, so that the extraction efficiency is influenced; according to the invention, the treatment cylinder can be kept in a totally closed state when a sample is cracked and mixed, so that the sample is prevented from leaking and being polluted; moreover, adsorption, washing purification and elution of nucleic acid can be sequentially completed, so that the whole process of nucleic acid extraction can be continuously completed, and the extraction efficiency is remarkably improved.

Description

Technical Field

[0001] This invention relates to the field of nucleic acid extraction technology, and in particular to a high-efficiency nucleic acid extraction adsorption column. Background Technology

[0002] With the widespread application of nucleic acid testing in clinical diagnosis, disease research, and forensic identification, accurate and efficient nucleic acid extraction is a crucial prerequisite for subsequent experiments. Currently, column-based nucleic acid extraction methods have become one of the mainstream methods due to their ease of operation and high recovery rate. This method typically uses a nucleic acid extraction adsorption column to extract nucleic acids from the sample. The adsorption column contains a lysis layer where lysis buffer is added to break cells and release nucleic acids, and an adsorption layer that adsorbs nucleic acids from the lysed mixture. The nucleic acids on the adsorption layer are then washed and purified, and finally eluted with an elution buffer to complete the nucleic acid extraction process.

[0003] With the development of technology, technicians in related fields have also optimized nucleic acid extraction adsorption columns accordingly. For example, Chinese patent CN110437980A discloses a nucleic acid extraction adsorption column, including an outer tube of the adsorption column, a filter head installed at the lower end of the outer tube, the lower end of the adsorption column being movable up and down relative to its outer tube, a sealing sleeve on the adsorption column, and an adsorption hole for nucleic acid to pass through at the lower end of the adsorption column; it also includes a pipette, which includes a pipette with an opening at the upper end, and a sealing head at the lower end of the pipette that can block the adsorption hole; in use, the nucleic acid solution is added to the pipette and remains in the pipette under the action of the sealing head blocking the adsorption column, and then the pipette and the sealing head are pulled upward to release the blockage of the adsorption hole, and the nucleic acid solution enters the filter head and the outer tube of the adsorption column in sequence through the pipette hole and the adsorption hole, and then the nucleic acid solution is discharged through the filter head to complete the extraction adsorption process.

[0004] However, the aforementioned existing technologies still have some shortcomings in the process of nucleic acid extraction: 1. Because existing technology only achieves partial sealing through the sealing head, it cannot completely seal the pipette. As a result, the pipette seal is easily lost during the mixing process of sample and lysis buffer due to shaking, centrifugation, etc., causing leakage of sample and lysis buffer. In addition, impurities in the air can easily enter the pipette and contaminate the sample, affecting the purity of nucleic acid.

[0005] 2. It can only complete the initial adsorption of nucleic acids. The adsorption column needs to be transferred to other containers for subsequent washing, purification and elution collection. This not only increases the number of operation steps and the risk of contamination during the transfer process, but also easily leads to the loss of nucleic acids during the transfer, affecting the extraction efficiency. In addition, when adsorbing nucleic acids, it is not possible to fully adsorb the nucleic acids in the solution by only using the filter head to adsorb them once. As a result, the nucleic acids are easy to flow out with the solution, resulting in low purity of the final extracted nucleic acids.

[0006] Therefore, based on the above-mentioned viewpoints, there is still room for improvement in existing nucleic acid extraction methods. Summary of the Invention

[0007] To address the aforementioned issues, this invention provides a high-efficiency nucleic acid extraction adsorption column, comprising: a processing tube with a top cap snapped onto its upper end; a mixing and lysis unit installed inside the processing tube, used to fully mix the nucleic acid sample and lysis buffer after the processing tube is completely sealed, releasing nucleic acid through cell lysis; an extraction tube detachably fitted onto the outer wall of the processing tube, with a silica gel membrane inside for adsorbing nucleic acid and intercepting the mixture flowing out of the processing tube, thus achieving nucleic acid adsorption, extraction, washing, and purification; a release unit installed between the processing tube and the extraction tube, used to discharge the lysed mixture from the processing tube into the extraction tube for nucleic acid extraction; and an elution tube detachably fitted onto the bottom of the extraction tube, with a waste liquid tube or a receiving tube detachably connected to the bottom of the elution tube, the receiving tube being used to collect the washed and purified nucleic acid.

[0008] As a preferred technical solution of the present invention, the mixing and pyrolysis unit includes a horizontal plate fixedly installed on the inner wall of the processing cylinder. A perforation is opened in the middle of the horizontal plate, and two elastic rubber sheets are symmetrically arranged in the perforation. The opposite sides of the two elastic rubber sheets abut against each other vertically. A positioning ring is also fixedly installed on the inner wall of the processing cylinder. An annular groove is formed on the inner side wall of the positioning ring. An annular frame is slidably connected inside the annular groove. An elastic rubber ring is set at the bottom of the annular frame. A conical cylinder is set on the inner side wall of the annular frame. A stopper is set at the bottom of the conical cylinder. A feeding hole connected to the conical cylinder is opened in the middle of the stopper. The thickness of the stopper gradually decreases from top to bottom, and the stopper is located above the gap between two elastic rubber sheets. As a preferred embodiment of the present invention, a plurality of annularly distributed stirring racks are uniformly installed between the lower end of the horizontal plate and the bottom wall of the processing cylinder, and a plurality of positioning strips are uniformly arranged on the bottom wall of the processing cylinder in an alternating pattern with the stirring racks.

[0009] As a preferred technical solution of the present invention, the release unit includes a backing plate installed on the inner wall of the extraction cylinder, the backing plate abutting against the bottom of the processing cylinder, both the bottom of the processing cylinder and the backing plate are tapered structures with diameters gradually decreasing from top to bottom, multiple sets of annularly distributed drainage holes are opened in the middle of the lower end of the processing cylinder, and multiple through holes corresponding to the positions of the drainage holes are opened on the backing plate. Multiple positioning blocks are installed on the upper part of the abutment plate, which are staggered with the through holes. A rubber plug is provided on the upper part of the positioning block to abut against the bottom of the drain hole.

[0010] As a preferred technical solution of the present invention, a plurality of air holes corresponding to the positions of the drain holes are evenly opened on the horizontal plate, and a plurality of connecting holes corresponding to the positions of the air holes are opened on the outer wall of the treatment cylinder. The connecting holes are located above the horizontal plate, and a connecting pipe is installed between the air holes and the connecting holes. A filter pad is provided at the upper end of the air holes. The upper end of the extraction cylinder is equipped with multiple support rods corresponding to the positions of the connecting holes. A rubber ball that slides against the outer wall of the connecting hole is installed on the side of the support rods near the processing cylinder.

[0011] As a preferred embodiment of the present invention, two silica gel membranes are installed on the inner wall of the extraction cylinder, one above the other. The upper silica gel membrane is used for coarse filtration of the mixture, and the lower silica gel membrane is used for fine filtration of the mixture. The silica gel membranes have an arc-shaped structure.

[0012] As a preferred embodiment of the present invention, the bottom of the extraction cylinder is detachably connected to a primary cylinder and a secondary cylinder in sequence, and a sealing ring is installed at the joint of the extraction cylinder, the primary cylinder and the secondary cylinder. Both the extraction cylinder and the first-stage cylinder have retaining rings installed on their inner walls at the top of the silicone membrane. Both the first-stage and second-stage cylinders have perforated plates installed on their inner walls to work with the retaining rings to clamp and fix the silicone membrane.

[0013] As a preferred embodiment of the present invention, a baffle is provided at the bottom of the secondary cylinder, and a plurality of annularly distributed discharge holes are evenly provided on the baffle. The washing cylinder is detachably sleeved on the outer wall of the secondary cylinder. A sealing plate is installed on the inner wall of the washing cylinder and rotates against the lower end of the baffle. A plurality of connecting holes corresponding to the positions of the discharge holes are provided on the sealing plate, and a plurality of sealing blocks are installed at the upper end of the sealing plate, which are staggered with the connecting holes. The sealing blocks slide against the bottom of the discharge holes.

[0014] As a preferred embodiment of the present invention, two fixing rubber rings are fitted on the bottom of the outer wall of both the processing cylinder and the secondary cylinder, and annular grooves that cooperate with the fixing rubber rings are opened on the inner wall of both the extraction cylinder and the elution cylinder.

[0015] As a preferred embodiment of the present invention, the upper half of the elution cylinder is a tapered structure with a gradually decreasing diameter, and the lower half of the elution cylinder is composed of multiple connecting cylinders with decreasing diameters, and the outer walls of the multiple connecting cylinders are fitted with auxiliary rubber rings.

[0016] In summary, this application includes the following beneficial technical effects: I. This invention, through the cooperation between the mixing and lysis unit and the release unit, can maintain the fully enclosed state of the processing tube during sample lysis and mixing, avoiding sample leakage and contamination; through the detachable connection and sealed cooperation between the extraction tube and the elution tube, nucleic acid adsorption, washing purification and elution can be completed sequentially, so that the entire process of nucleic acid extraction can be completed continuously with only one adsorption column, which significantly improves the convenience of operation and extraction efficiency.

[0017] Second, this invention can seal the upper and lower ends of the processing tube respectively through the elastic rubber sheet and the release unit, ensuring that the lysis buffer and nucleic acid sample can be inverted and shaken, shaken up and down, or placed on a centrifuge for centrifugation in the fully sealed processing tube without overflowing during the mixing process. In addition, the mixing method of shaking and centrifugation can avoid the deposition of high concentration salt ions in the lysis buffer at the bottom of the sample, which may lead to local over-lysis or local incomplete lysis. This achieves rapid mixing of lysis buffer and nucleic acid sample, ensures uniform mixing, and improves the lysis efficiency of nucleic acid.

[0018] Third, this invention enables the processing tube to be fully sealed when the nucleic acid sample and lysis buffer are shaken or centrifuged by conveniently controlling the sealing and opening of the upper and lower ends of the processing tube, thus avoiding leakage of the mixture and ensuring sufficient mixing between the nucleic acid sample and the lysis buffer. The convenient opening of the upper and lower ends of the processing tube allows the mixed mixture to flow smoothly downward into the extraction tube for nucleic acid adsorption and extraction.

[0019] Fourth, this invention enables dual filtration of the mixture using two silicone membranes, which can effectively adsorb nucleic acids in the mixture. The silicone membranes have an arc-shaped structure, which increases the contact area with the mixture, thereby improving the filtration effect and facilitating the uniform distribution of the mixture, thus avoiding local blockage. Attached Figure Description

[0020] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0021] Figure 1 This is a schematic diagram of the structure of the present invention.

[0022] Figure 2 This is a schematic diagram of the structure of the hybrid pyrolysis unit of the present invention.

[0023] Figure 3 This is the present invention. Figure 2 A magnified view of part A.

[0024] Figure 4 This is the present invention. Figure 2 A magnified view of section B.

[0025] Figure 5 This is a schematic diagram of the structure between the processing cylinder, extraction cylinder and release unit of the present invention.

[0026] Figure 6 This is the present invention. Figure 5 A magnified view of a portion of point C.

[0027] Figure 7 This is a schematic diagram of the structure between the drain hole, through hole and positioning block of the present invention.

[0028] Figure 8 This is the present invention. Figure 5 A magnified view of a portion of point D.

[0029] Figure 9 This is a schematic diagram of the internal structure between the extraction cylinder, the primary cylinder, and the secondary cylinder of the present invention.

[0030] Figure 10 This is a schematic diagram of the exploded structure between the extraction cylinder, the primary cylinder, and the secondary cylinder of the present invention.

[0031] In the diagram, 1. Processing cylinder; 11. Top cover; 12. Fixing rubber ring; 2. Mixing and pyrolysis unit; 21. Horizontal plate; 22. Elastic rubber sheet; 23. Positioning ring; 24. Ring frame; 25. Elastic rubber ring; 26. Conical cylinder; 27. Plug; 28. Discharge hole; 29. ​​Stirring frame; 20. Positioning strip; 3. Extraction cylinder; 31. Silica gel membrane; 32. Primary cylinder; 33. Secondary cylinder; 331. Baffle; 332. Discharge hole; 333. Sealing plate; 334. Connecting hole; 335. Sealing block; 34. Fixing retaining ring; 35. Mesh plate; 4. Release unit; 41. Abutment plate; 42. Drain hole; 43. Through hole; 44. Positioning block; 45. Rubber plug; 46. Connecting hole; 47. Filter pad; 48. Support rod; 49. Rubber ball; 5. Eluent cylinder; 51. Receiving cylinder. Detailed Implementation

[0032] The following is in conjunction with the appendix Figures 1-10 The embodiments of the present invention will be described in detail below.

[0033] This application discloses a high-efficiency nucleic acid extraction adsorption column. It should be noted that the high-efficiency nucleic acid extraction adsorption column of this application is mainly used in the process of nucleic acid extraction. In terms of technical effect, it can maintain the fully sealed state of the processing tube 1 when the sample is lysed and mixed, avoiding leakage and contamination of the mixture formed by the nucleic acid sample and the lysis solution. Furthermore, the high-efficiency nucleic acid extraction adsorption column of this application can also sequentially complete the adsorption, washing purification and elution of nucleic acid, so that the entire process of nucleic acid extraction can be completed continuously with only one adsorption column, which significantly improves the extraction efficiency.

[0034] Reference Figure 1As shown, a high-efficiency nucleic acid extraction adsorption column includes a processing tube 1 with a top cap 11 snapped onto its upper end; a mixing and lysis unit 2 installed inside the processing tube 1 to fully seal the processing tube 1 and thoroughly mix the nucleic acid sample and lysis buffer, releasing nucleic acid through cell lysis in the sample; an extraction tube 3 detachably fitted onto the outer wall of the processing tube 1, with a silica gel membrane 31 inside the extraction tube 3 for adsorbing nucleic acid and intercepting the mixture flowing out of the processing tube 1, thus achieving nucleic acid adsorption, extraction, washing, and purification; a release unit 4 installed between the processing tube 1 and the extraction tube 3 to discharge the lysed mixture from the processing tube 1 into the extraction tube 3 for nucleic acid extraction; through the cooperation of the mixing and lysis unit 2 and the release unit 4, the processing tube 1 is kept fully sealed during mixing of the sample and lysis buffer, preventing sample outflow; and an elution tube 5 detachably fitted onto the bottom of the extraction tube 3, with a waste liquid tube or a receiving tube 51 detachably connected to the bottom of the elution tube 5, which is used to collect and wash the purified nucleic acid.

[0035] In the specific implementation process, firstly, the nucleic acid sample is added to the processing tube 1 through the sample loading chamber. Secondly, lysis buffer is added to the processing tube 1. Then, the sample and lysis buffer are thoroughly mixed by inverting, shaking, and centrifuging, so that the cells inside the sample lyse and release nucleic acid. At this time, the sample and lysis buffer form a mixture containing nucleic acid. During this period, the release unit 4 ensures that the processing tube 1 remains in a completely sealed state, preventing the mixture from flowing out when shaking or centrifuging the processing tube 1. It also works with the mixing and lysis unit 2 to thoroughly mix the sample and lysis buffer. Then, the bottom of the processing tube 1 is opened through the release unit 4, and the present invention is placed... On a centrifuge, while the centrifuge controls the rotation of this invention, the mixture in the processing tube 1 flows downward into the extraction tube 3 and passes through the silica membrane 31 under centrifugal action, which is used to adsorb the nucleic acid in the mixture onto the silica membrane 31. Then, the invention is removed from the centrifuge, and washing solution is added to the processing tube 1. The washing solution flows downward after passing through the processing tube 1 and washes and purifies the nucleic acid adsorbed on the silica membrane 31, thereby removing impurities, while the nucleic acid is still adsorbed on the silica membrane 31. It should be noted that during nucleic acid washing and purification, the bottom of the elution tube 5 is connected to the waste liquid pipe, and the washing solution carrying impurities flows out into the waste liquid pipe after passing through the silica membrane 31.

[0036] Next, the lower end of the extraction cylinder 3 is sealed by the elution cylinder 5, and then the eluent is added to the extraction cylinder 3. The upper end of the extraction cylinder 3 is then sealed by the release unit 4. The present invention is then placed back on the centrifuge for centrifugation and rotation, so that the eluent is in full contact with the silica membrane 31 to elute the purified nucleic acid on the silica membrane 31. Finally, the receiving cylinder 51 is installed at the bottom of the elution cylinder 5, and the extraction cylinder 3 is opened by the elution cylinder 5, so that the eluent carrying the nucleic acid flows out into the receiving cylinder 51, thus completing the extraction of nucleic acid.

[0037] Reference Figure 2 , Figure 3 and Figure 4 As shown, in order to improve the lysis efficiency of the sample and avoid incomplete lysis, a hybrid lysis unit 2 is provided in this embodiment. Specifically, the hybrid lysis unit 2 includes a horizontal plate 21 fixedly installed on the inner wall of the processing cylinder 1. A perforation is opened in the middle of the horizontal plate 21, and two elastic rubber sheets 22 are symmetrically arranged in the perforation. The opposite sides of the two elastic rubber sheets 22 abut against each other vertically. A positioning ring 23 is also fixedly installed on the inner wall of the processing cylinder 1. An annular groove is opened on the inner side wall of the positioning ring 23. An annular frame 24 is slidably connected inside the annular groove. An elastic rubber ring 25 is provided at the bottom of the annular frame 24. A conical cylinder 26 is provided on the inner side wall of the annular frame 24. A plug 27 is provided at the bottom of the conical cylinder 26. A feeding hole 28 communicating with the conical cylinder 26 is opened in the middle of the plug 27. The thickness of the plug 27 gradually decreases from top to bottom, and the plug 27 is located above the gap between the two elastic rubber sheets 22. It should be noted that both the elastic rubber sheet 22 and the elastic rubber ring 25 are elastic materials and can undergo elastic deformation under external force. Therefore, the ring frame 24 moves upward in the initial state under the elastic action of the elastic rubber ring 25, and the ring frame 24 drives the plug 27 to move upward to the highest position through the conical cylinder 26. At this time, the plug 27 is located above the elastic rubber sheet 22, so that the two elastic rubber sheets 22 abut against each other to seal the upper end of the processing cylinder 1 and prevent the processing cylinder 1 from communicating with the outside and allowing impurities to enter.

[0038] Furthermore, in this embodiment, a plurality of annularly distributed stirring racks 29 are uniformly installed between the lower end of the horizontal plate 21 and the inner bottom wall of the processing cylinder 1, and a plurality of positioning strips 20 are uniformly arranged on the inner bottom wall of the processing cylinder 1, which are staggered with the stirring racks 29.

[0039] In the specific implementation process, when it is necessary to inject lysis buffer and nucleic acid samples into the processing cylinder 1, the outlet of the sample loading chamber is pressed against the conical cylinder 26, and then the conical cylinder 26 is pressed down through the sample loading chamber. The conical cylinder 26 drives the stopper 27 to pass through the gap between the two elastic rubber sheets 22, so that the discharge hole 28 is connected to the processing cylinder 1, so that the sample loading chamber can add the lysis buffer and nucleic acid samples into the processing cylinder 1 in sequence through the discharge hole 28. After the lysis buffer and nucleic acid samples have been added, the sample loading chamber is removed from the conical cylinder 26. Under the action of the elastic rubber ring 25, the conical cylinder 26 drives the stopper 27 to move upward and reset, and the two elastic rubber sheets 22 resume contact, thereby restoring the seal on the upper end of the processing cylinder 1.

[0040] The lysis buffer and nucleic acid sample are then inverted and shaken, or placed in a centrifuge for centrifugation to form a mixture. During this process, the stirring rack 29 and positioning bar 20 move relative to the mixture to assist in rapid mixing and prevent high concentrations of salt ions in the lysis buffer from depositing at the bottom of the sample, which could lead to local over-lysis or incomplete lysis. This ensures rapid mixing of the lysis buffer and nucleic acid sample, guarantees uniform mixing, and improves nucleic acid lysis efficiency. It should be noted that since both the top and bottom of the processing cylinder 1 are sealed, shaking and centrifuging the mixture in the fully enclosed processing cylinder 1 effectively prevents overflow.

[0041] Reference Figure 5 , Figure 6 and Figure 7 As shown, in order to facilitate the sealing of the bottom of the processing tube 1 and to ensure that the nucleic acid sample and lysis buffer are fully mixed in a completely sealed state, the bottom of the processing tube 1 can be sealed by the extraction tube 3 in this embodiment. Based on this, this application provides a release unit 4, which includes a contact plate 41 installed on the inner wall of the extraction tube 3. The contact plate 41 abuts against the bottom of the processing tube 1. Both the bottom of the processing tube 1 and the contact plate 41 are conical structures with a diameter that gradually decreases from top to bottom. Multiple sets of annularly distributed drainage holes 42 are opened in the middle of the lower end of the processing tube 1. Multiple through holes 43 corresponding to the positions of the drainage holes 42 are opened on the contact plate 41. Multiple positioning blocks 44 are installed on the upper end of the contact plate 41, which are staggered with the through holes 43. A rubber stopper 45 abuts against the bottom of the drainage hole 42 on the upper end of the positioning block 44.

[0042] In the initial state, the positioning block 44 drives the rubber stopper 45 to abut against the bottom of the drain hole 42 to seal the drain hole 42, thereby sealing the bottom of the processing cylinder 1. At this time, it is convenient to fully mix the nucleic acid sample and lysis buffer by shaking or centrifugal rotation.

[0043] In the specific implementation process, after the nucleic acid sample and lysis buffer are mixed, a mixture containing nucleic acid and impurities is formed. Then, the processing cylinder 1 and the extraction cylinder 3 are rotated relative to each other, so that the extraction cylinder 3 moves the rubber stopper 45 away from the drain hole 42 through the positioning block 44, and the control hole 43 is connected to the drain hole 42, thereby releasing the seal on the bottom of the processing cylinder 1 and connecting the processing cylinder 1 and the extraction cylinder 3. Subsequently, the present invention is placed on a centrifuge for rotation. Under the centrifugation action, the mixture in the processing cylinder 1 passes through the drain hole 42 and the through hole 43 and enters the extraction cylinder 3, and passes through the silica membrane 31 inside the extraction cylinder 3 to filter out impurities, and the nucleic acid in the mixture is adsorbed on the silica membrane 31.

[0044] Reference Figure 5 and Figure 8As shown, when no nucleic acid sample and lysis buffer are injected, the upper end of the processing cylinder 1 remains closed. Therefore, it is easy for a negative pressure to form inside the processing cylinder 1 after the drain hole 42 is opened, which prevents the mixture from flowing out smoothly. Based on this, in this embodiment, multiple air holes corresponding to the positions of the drain hole 42 are evenly opened on the horizontal plate 21. Multiple connecting holes 46 corresponding to the positions of the air holes are opened on the outer wall of the processing cylinder 1. The connecting holes 46 are located above the horizontal plate 21. A connecting pipe is installed between the air holes and the connecting holes 46. A filter pad 47 is provided at the upper end of the air holes. Multiple support rods 48 corresponding to the positions of the connecting holes 46 are provided at the upper end of the extraction cylinder 3. A rubber ball 49 that slides against the outer wall of the connecting hole 46 is installed on the side of the support rod 48 near the processing cylinder 1.

[0045] In the initial state, the support rod 48 drives the rubber ball 49 to abut against the connecting hole 46 to seal the connecting hole 46, thereby preventing the processing cylinder 1 from communicating with the outside and disrupting the closed state of the processing cylinder 1; in addition, when the processing cylinder 1 is shaken or centrifuged, the filter pad 47 can block the mixture, preventing the mixture from overflowing upward through the pores, and the filter pad 47 is preferably a sterile medical degreased cotton ball, so that even if the mixture comes into contact with the filter pad 47, the purity of the nucleic acid can be guaranteed not to be affected.

[0046] In the specific implementation process, after the nucleic acid sample and lysis buffer are mixed, the processing cylinder 1 and the extraction cylinder 3 rotate relative to each other. At the same time, the drain hole 42 is connected to the through hole 43, and the processing cylinder 1 and the multiple support rods 48 rotate relative to each other, so that the rubber ball 49 is misaligned with the connecting hole 46. Thus, the vent is connected to the outside through the connecting tube and the connecting hole 46. When the present invention is centrifugally rotated, the outside air can enter the interior of the processing cylinder 1 through the connecting hole 46, avoiding the negative pressure inside the processing cylinder 1 from affecting the downward flow of the mixed liquid. In addition, the filter pad 47 can also filter the air entering the processing cylinder 1 to intercept dust particles, bacteria and other pollutants in the air, and prevent them from being mixed into the mixed liquid with the air and contaminating the nucleic acid sample, thus affecting the final nucleic acid purity.

[0047] Reference Figure 9 and Figure 10 As shown, in order to enhance the extraction effect of nucleic acid in the mixture, in this embodiment, two silica gel membranes 31 are installed on the inner wall of the extraction cylinder 3. The upper silica gel membrane 31 is used for coarse filtration of the mixture, and the lower silica gel membrane 31 is used for fine filtration of the mixture. This can effectively intercept and adsorb nucleic acid in the mixture, and prevent a small amount of nucleic acid from flowing out after passing through the silica gel membrane 31 with impurities.

[0048] Furthermore, in this embodiment, the bottom of the extraction cylinder 3 is detachably connected to a primary cylinder 32 and a secondary cylinder 33 in sequence. Sealing rings are installed at the joints of the extraction cylinder 3, the primary cylinder 32, and the secondary cylinder 33 to ensure that the interior of the extraction cylinder 3 remains sealed and to prevent the mixture from overflowing during adsorption and subsequent elution. The inner walls of the extraction cylinder 3 and the primary cylinder 32 are each equipped with a fixing ring 34 located at the upper end of the silicone membrane 31. The inner walls of the primary cylinder 32 and the secondary cylinder 33 are each equipped with a perforated plate 35, which is used to cooperate with the fixing ring 34 to clamp and fix the silicone membrane 31.

[0049] In the specific implementation process, when installing the extraction cylinder 3, the coarsely filtered silica membrane 31 is first placed on the upper end of the mesh plate 35 on the inner wall of the primary cylinder 32, and then the primary cylinder 32 is inserted into the bottom of the extraction cylinder 3. The mesh plate 35 on the inner wall of the primary cylinder 32 abuts against the fixing ring 34 on the inner wall of the extraction cylinder 3, thus fixing the silica membrane 31.

[0050] Next, the finely filtered silica membrane 31 is placed on the upper end of the mesh plate 35 on the inner wall of the secondary cylinder 33. Then, the secondary cylinder 33 is inserted into the bottom of the primary cylinder 32, and the silica membrane 31 is fixed by the fixing ring 34 and the mesh plate 35 between the primary cylinder 32 and the secondary cylinder 33. This completes the quick installation of the extraction cylinder 3, the primary cylinder 32, the secondary cylinder 33 and the silica membrane 31.

[0051] When the mixture is adsorbed and filtered, the mixture can be filtered twice through two silica membranes 31, which can effectively adsorb nucleic acids in the mixture. The silica membrane 31 has an arc-shaped structure, which increases the contact area with the mixture, improves the filtration effect, and facilitates the uniform distribution of the mixture, avoiding local blockage.

[0052] Continue to refer to Figure 9 and Figure 10 As shown, in order to facilitate the complete elution of nucleic acids on the silica membrane 31, in this embodiment, a baffle 331 is provided at the bottom of the secondary cylinder 33. The baffle 331 has a plurality of annularly distributed discharge holes 332 evenly distributed on it. The elution cylinder 5 is detachably sleeved on the outer wall of the secondary cylinder 33. A sealing plate 333 is installed on the inner wall of the elution cylinder 5 and rotates against the lower end of the baffle 331. The sealing plate 333 has a plurality of connecting holes 334 corresponding to the positions of the discharge holes 332. A plurality of sealing blocks 335 are installed on the upper end of the sealing plate 333 and are staggered with the connecting holes 334. The sealing blocks 335 slide against the bottom of the discharge holes 332.

[0053] In the initial state, the sealing block 335 abuts against the lower end of the discharge hole 332 to block the discharge hole 332, thereby sealing the bottom of the extraction cylinder 3 so that the mixture will not continue to flow downward when it flows downward into the extraction cylinder 3.

[0054] "In the specific implementation process, after the mixture in the processing cylinder 1 is completely flowed into the extraction cylinder 3 by centrifugal rotation, the processing cylinder 1 and the extraction cylinder 3 are rotated relative to each other, so that the extraction cylinder 3 drives the rubber stopper 45 to restore the seal on the drain hole 42 through the positioning block 44, thereby achieving the sealing of the upper and lower ends of the extraction cylinder 3; then the present invention is placed on a centrifuge to centrifuge the mixture, and under the centrifugal action, the mixture in the extraction cylinder 3 flows downward and passes through two silica membranes 31 in sequence for double filtration of the mixture, so that the nucleic acid in the mixture is adsorbed on the silica membrane 31, and the filtered mixture accumulates at the bottom of the baffle 331 after passing through the silica membrane 31."

[0055] After the mixed liquid adsorption and filtration is completed, the elution cylinder 5 is connected to the waste liquid pipe. Then, the treatment cylinder 1 and the extraction cylinder 3 are rotated relative to each other, so that the rubber stopper 45 is misaligned with the drain hole 42, and the seal at the upper end of the extraction cylinder 3 is released. Subsequently, the elution cylinder 5 and the secondary cylinder 33 are rotated relative to each other, so that the sealing block 335 is misaligned with the discharge hole 332, and the seal at the lower end of the extraction cylinder 3 is released. The mixed liquid flows into the waste liquid pipe after passing through the discharge hole 332 and the elution cylinder 5.

[0056] Subsequently, washing solution is added to the processing cylinder 1. The washing solution is used to remove impurities such as proteins and polysaccharides adsorbed on the silica membrane 31. At the same time, it provides a high-salt, slightly acidic, or neutral pH environment to ensure that nucleic acids can be firmly adsorbed on the surface of the silica membrane 31. After the washing solution passes through the drain hole 42, it enters the extraction cylinder 3, where it temporarily remains on the silica membrane 31. Then, the top and bottom ends of the extraction cylinder 3 are sealed again, and the invention is placed on a centrifuge for rotation, thereby extending the contact time between the washing solution and the silica membrane 31. This allows for thorough washing and purification of the mixture on the silica membrane 31. The washing solution carries impurities downwards to the baffle 331, while the nucleic acids remain adsorbed on the silica membrane 31. Centrifugal rotation allows the washing solution to pass through the silica membrane 31 quickly and evenly, carrying impurities from the surface and pores of the silica membrane 31 downwards. This effectively improves the separation efficiency of impurities and nucleic acids and avoids impurity residue affecting the purity of nucleic acids.

[0057] Next, the seals at both ends of the extraction cylinder 3 are released, and the washing liquid containing impurities is discharged into the waste liquid pipe. Then, eluent is added to the processing cylinder 1. The eluent is used to create a low-salt, neutral, or weakly alkaline pH environment, causing the nucleic acid to dissociate from the silica membrane 31. The eluent passes through the drain hole 42 and enters the extraction cylinder 3. The upper and lower ends of the extraction cylinder 3 are sealed again. Then, the invention is placed on a centrifuge and centrifuged to allow the eluent to quickly and completely wet the silica membrane 31, accelerating the dissociation of nucleic acid from the silica membrane 31. The eluent carries the nucleic acid downwards and accumulates at the upper end of the baffle 331. Then, the elution cylinder 5 is connected to the receiving cylinder 51. Finally, the seals at both ends of the extraction cylinder 3 are released, allowing the eluent carrying the nucleic acid to flow into the receiving cylinder 51, thereby achieving the lysis, purification, and extraction of nucleic acid. Continue to refer to Figure 9 and Figure 10 As shown, in order to facilitate the quick installation between the processing cylinder 1 and the extraction cylinder 3, and the elution cylinder 5 and the secondary cylinder 33, and to achieve the sealing of the processing cylinder 1 and the extraction cylinder 3 by rotation after installation, in this embodiment, two fixing rubber rings 12 are fitted on the bottom of the outer wall of the processing cylinder 1 and the secondary cylinder 33, and the inner wall of the extraction cylinder 3 and the elution cylinder 5 are provided with annular grooves that cooperate with the fixing rubber rings 12. The cooperation between the fixing rubber rings 12 and the annular grooves facilitates the quick installation and disassembly of the processing cylinder 1 and the extraction cylinder 3, and the elution cylinder 5 and the secondary cylinder 33. While maintaining a sealed environment, it can also ensure that the processing cylinder 1 and the extraction cylinder 3, and the elution cylinder 5 and the secondary cylinder 33 can rotate relative to each other.

[0058] Reference Figure 10 As shown, in order to facilitate the adaptation of waste liquid pipes and receiving cylinders 51 of different diameters at the bottom of the washing cylinder 5, in this embodiment, the upper half of the washing cylinder 5 is a tapered structure with a gradually decreasing diameter, and the lower half of the washing cylinder 5 is composed of multiple connecting cylinders with decreasing diameters, which are used to adapt to waste liquid pipes and receiving cylinders 51 of different diameters. The outer walls of the multiple connecting cylinders are fitted with auxiliary rubber rings, which are used to fix the waste liquid pipes and receiving cylinders 51 accordingly, and can seal the installed waste liquid pipes or receiving cylinders 51 to prevent leakage, and facilitate the quick installation and disassembly of waste liquid pipes and receiving cylinders 51.

[0059] During operation: Step 1: First, place the outlet of the sample loading chamber against the conical cylinder 26, and then press the conical cylinder 26 downward through the sample loading chamber. The conical cylinder 26 drives the stopper 27 through the gap between the two elastic rubber sheets 22 and connects with the processing cylinder 1, so that the sample loading chamber can sequentially add the lysis buffer and nucleic acid sample into the processing cylinder 1 through the discharge hole 28. After both the lysis buffer and nucleic acid sample have been added, remove the sample loading chamber from the conical cylinder 26. Under the action of the elastic rubber ring 25, the conical cylinder 26 drives the stopper 27 to move upward and reset. The two elastic rubber sheets 22 resume contact and seal the upper end of the processing cylinder 1. Then, the present invention, carrying the lysis buffer and nucleic acid sample, is inverted and shaken, shaken up and down, or placed on a centrifuge for centrifugal rotation to form a mixture.

[0060] Step 2: Rotate the processing cylinder 1 and the extraction cylinder 3 relative to each other. The extraction cylinder 3 moves the rubber stopper 45 away from the drain hole 42 through the positioning block 44, releasing the seal on the bottom of the processing cylinder 1 and connecting the processing cylinder 1 and the extraction cylinder 3. Then, place the present invention on a centrifuge and rotate it. Under the centrifugal action, the mixture in the processing cylinder 1 passes through the drain hole 42 and the through hole 43 and enters the extraction cylinder 3.

[0061] Step 3: Rotate the processing cylinder 1 and the extraction cylinder 3 relative to each other, so that the extraction cylinder 3, through the positioning block 44, drives the rubber stopper 45 to restore the seal on the drain hole 42, and cooperates with the baffle 331 to seal the upper and lower ends of the extraction cylinder 3; then place the present invention on a centrifuge to centrifuge the mixture. Under the centrifugation action, the mixture in the extraction cylinder 3 flows downward and passes through two silica membranes 31 in sequence for double filtration of the mixture, so that the nucleic acid in the mixture is adsorbed on the silica membrane 31, and the filtered mixture accumulates at the bottom of the baffle 331 after passing through the silica membrane 31.

[0062] After the adsorption and filtration of the mixed liquid is completed, the seals at both ends of the extraction cylinder 3 are released, and the mixed liquid flows into the waste liquid pipe after passing through the discharge hole 332 and the washing cylinder 5.

[0063] Step 4: Add washing solution to processing cylinder 1. The washing solution enters extraction cylinder 3 after passing through drainage hole 42, so that the washing solution is temporarily stored on silica membrane 31. Then restore the seal at both ends of extraction cylinder 3, and place the present invention on a centrifuge to rotate, so that the washing solution can fully wash and purify the mixture on silica membrane 31. The washing solution carries impurities such as proteins and polysaccharides and flows downward to baffle 331, while nucleic acids are still adsorbed on silica membrane 31.

[0064] Step 5: Unseal the top and bottom ends of extraction cylinder 3, drain the washing liquid containing impurities into the waste liquid pipe, then add eluent to processing cylinder 1. The eluent passes through the drain hole 42 and enters extraction cylinder 3. Seal the top and bottom ends of extraction cylinder 3 again, then place the present invention on a centrifuge for centrifugal rotation, so that the eluent can quickly and completely wet the silica membrane 31, accelerating the dissociation of nucleic acid from silica membrane 31. Finally, unseal the top and bottom ends of extraction cylinder 3, so that the eluent carries the nucleic acid into receiving cylinder 51, thereby realizing the lysis, purification and extraction of nucleic acid.

[0065] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the present invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the present invention.

[0066] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A high-efficiency nucleic acid extraction adsorption column, characterized in that, include: Processing cylinder (1), with a top cover (11) snapped onto the upper end of the processing cylinder (1); The mixing and lysis unit (2) is installed inside the processing tube (1) to fully mix the nucleic acid sample and lysis buffer after the processing tube (1) is completely sealed, and the cells in the sample are lysed to release nucleic acid; The extraction tube (3) is detachably sleeved on the outer wall of the processing tube (1). The extraction tube (3) is equipped with a silica membrane (31) for adsorbing nucleic acids, which is used to intercept the mixed liquid flowing out of the processing tube (1) to realize the adsorption extraction and washing purification of nucleic acids. The release unit (4) is installed between the processing tube (1) and the extraction tube (3) to discharge the lysed mixture in the processing tube (1) into the extraction tube (3) to extract nucleic acid. Through the cooperation of the mixing lysis unit (2) and the release unit (4), the sample and lysis solution are kept completely sealed in the processing tube (1) during mixing to prevent the sample from flowing out. The elution tube (5) is detachably attached to the bottom of the extraction tube (3). The bottom of the elution tube (5) is detachably connected to a waste liquid tube or a receiving tube (51). The receiving tube (51) is used to receive and collect the washed and purified nucleic acid.

2. The high-efficiency nucleic acid extraction adsorption column according to claim 1, characterized in that: The hybrid pyrolysis unit (2) includes a horizontal plate (21) fixedly installed on the inner wall of the processing cylinder (1). A perforation is provided in the middle of the horizontal plate (21), and two elastic rubber sheets (22) are symmetrically arranged in the perforation. The opposite sides of the two elastic rubber sheets (22) abut against each other vertically. A positioning ring (23) is fixedly installed on the inner wall of the processing cylinder (1). An annular groove is opened on the inner side wall of the positioning ring (23). An annular frame (24) is slidably connected inside the annular groove. An elastic rubber ring (25) is set at the bottom of the annular frame (24). A conical cylinder (26) is set on the inner side wall of the annular frame (24). A plug (27) is set at the bottom of the conical cylinder (26). A feeding hole (28) connected to the conical cylinder (26) is opened in the middle of the plug (27). The thickness of the plug (27) gradually decreases from top to bottom, and the plug (27) is located above the gap between two elastic rubber sheets (22).

3. The high-efficiency nucleic acid extraction adsorption column according to claim 2, characterized in that: Multiple ring-shaped stirring racks (29) are evenly installed between the lower end of the horizontal plate (21) and the inner bottom wall of the processing cylinder (1), and multiple positioning strips (20) are evenly arranged on the inner bottom wall of the processing cylinder (1) in an alternating manner with the stirring racks (29).

4. The high-efficiency nucleic acid extraction adsorption column according to claim 1, characterized in that: The release unit (4) includes a backing plate (41) installed on the inner wall of the extraction cylinder (3). The backing plate (41) abuts against the bottom of the processing cylinder (1). Both the bottom of the processing cylinder (1) and the backing plate (41) are conical structures with diameters gradually decreasing from top to bottom. Multiple sets of annularly distributed drainage holes (42) are opened in the middle of the lower end of the processing cylinder (1). Multiple through holes (43) corresponding to the positions of the drainage holes (42) are opened on the backing plate (41). The upper end of the abutment plate (41) is equipped with multiple positioning blocks (44) that are staggered with the through hole (43). The upper end of the positioning block (44) is provided with a rubber plug (45) that abuts against the bottom of the drain hole (42).

5. The high-efficiency nucleic acid extraction adsorption column according to claim 4, characterized in that: The horizontal plate (21) is evenly provided with a plurality of air holes corresponding to the position of the drain hole (42). The outer wall of the treatment cylinder (1) is provided with a plurality of connecting holes (46) corresponding to the position of the air holes. The connecting holes (46) are located above the horizontal plate (21). A connecting pipe is installed between the air holes and the connecting holes (46). A filter pad (47) is provided at the upper end of the air holes. The upper end of the extraction tube (3) is provided with multiple support rods (48) corresponding to the position of the connecting hole (46). A rubber ball (49) that slides against the outer wall of the connecting hole (46) is installed on the side of the support rod (48) near the processing tube (1).

6. The high-efficiency nucleic acid extraction adsorption column according to claim 1, characterized in that: The inner wall of the extraction tube (3) is fitted with two vertically distributed silica membranes (31). The upper silica membrane (31) is used for coarse filtration of the mixture, and the lower silica membrane (31) is used for fine filtration of the mixture. The silica membrane (31) has an arc-shaped structure.

7. The high-efficiency nucleic acid extraction adsorption column according to claim 1, characterized in that: The bottom of the extraction cylinder (3) is detachably connected to a primary cylinder (32) and a secondary cylinder (33). Sealing rings are installed at the joints of the extraction cylinder (3), the primary cylinder (32) and the secondary cylinder (33). The inner walls of the extraction tube (3) and the first-stage tube (32) are equipped with a fixing ring (34) located at the upper end of the silicone membrane (31). The inner walls of the first-stage tube (32) and the second-stage tube (33) are equipped with a mesh plate (35) to cooperate with the fixing ring (34) to clamp and fix the silicone membrane (31).

8. The high-efficiency nucleic acid extraction adsorption column according to claim 7, characterized in that: The bottom of the secondary cylinder (33) is provided with a baffle (331), and multiple annularly distributed discharge holes (332) are evenly opened on the baffle (331). The washing cylinder (5) is detachably sleeved on the outer wall of the secondary cylinder (33). The inner wall of the washing cylinder (5) is equipped with a sealing plate (333) that rotates and abuts against the lower end of the baffle (331). Multiple connecting holes (334) corresponding to the positions of the discharge holes (332) are opened on the sealing plate (333), and multiple sealing blocks (335) that are staggered with the connecting holes (334) are installed on the upper end of the sealing plate (333). The sealing blocks (335) slide against the bottom of the discharge holes (332).

9. The high-efficiency nucleic acid extraction adsorption column according to claim 7, characterized in that: The bottom of the outer wall of the processing cylinder (1) and the secondary cylinder (33) are fitted with two fixing rubber rings (12), and the inner wall of the extraction cylinder (3) and the elution cylinder (5) are provided with annular grooves that cooperate with the fixing rubber rings (12).

10. The high-efficiency nucleic acid extraction adsorption column according to claim 1, characterized in that: The upper half of the elution tube (5) is a tapered structure with a gradually decreasing diameter, and the lower half of the elution tube (5) is composed of multiple connecting tubes with decreasing diameters. The outer walls of the multiple connecting tubes are fitted with auxiliary rubber rings.

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