High-throughput gene splicing and construction workstation and gene splicing and construction method

Abstract

The invention discloses a high-throughput gene splicing and construction workstation and a gene splicing and construction method, and belongs to the technical field of gene engineering. Comprising a frame body, a moving module arranged on the frame body, and an enzyme label concentration detection module, an electrophoresis module, a suction filtration and purification module, a storage module capable of storing a preset solvent, a centrifugation module, a constant-temperature oscillation module, a film sealing module and a PCR instrument which can be in butt joint with the moving module, the rubber cutting module can be in butt joint with the storage module, and the monitoring assembly can be electrically connected with the above structures. According to preset information, primer preparation, gene splicing reaction liquid preparation, splicing reaction operation, gene product amplification reaction liquid preparation, amplification reaction operation, agarose gel electrophoresis, target gene strip cutting, electrophoresis PCR product purification, enzyme-labeled quantification and uniform concentration, enzyme linked reaction liquid preparation and enzyme linked reaction operation are carried out; the mobile module is provided with a liquid suction head for taking and placing the solvent, so that the flexibility and the applicability are good.

Description

Technical Field

[0001] The present invention relates to the technical field of gene engineering, and in particular to a high-throughput gene splicing and construction workstation and a gene splicing and construction method. Background Art

[0002] Automated gene splicing instruments, such as the iBioFAB system, can automatically complete the production of transcription activator-like effector nucleases (TALENs), reducing dependence on manual labor and improving overall experimental efficiency. However, most of the automation systems in the prior art focus on the automation of experimental operations, with large application limitations, making it difficult to achieve multi-purpose use of one machine, and still limited support for user customization needs and rapid response capabilities. There is an urgent need for a high-throughput gene splicing and construction workstation and a gene splicing and construction method to solve the above problems. Summary of the invention

[0003] The purpose of the present invention is to provide an automated workstation for high-throughput gene splicing and a gene splicing and construction method, so as to improve the flexibility, applicability and scalability of the workstation.

[0004] To achieve this object, the present invention adopts the following technical solutions:

[0005] A high-throughput gene splicing and construction workstation comprises a frame, on which are provided: a mobile module and an enzyme-labeled concentration detection module, an electrophoresis module, a filtration purification module, a storage module capable of storing preset solvents, a centrifugation module, a constant temperature oscillation module, a film sealing module and a PCR instrument, which can be docked with the mobile module respectively; a gel cutting module and a monitoring component are also provided on the frame, the gel cutting module can be docked with the storage module, the monitoring component can be electrically connected to the above structures respectively, and configure primers, configure gene splicing reaction liquid, run splicing reaction, configure gene product amplification reaction liquid, run amplification reaction, perform agarose gel electrophoresis, cut target gene bands, purify electrophoresis PCR products, quantify enzyme labels and make concentration uniform, configure enzyme-linked reaction liquid and run enzyme-linked reaction according to preset information; the mobile module is equipped with a liquid pipette head for taking and placing solvents.

[0006] In some possible embodiments, the mobile module includes a first mobile component, a second mobile component, a 96-channel pipetting component, an 8-channel pipetting component and a carrying clamp, all of which are electrically connected to the monitoring component, the 96-channel pipetting component is connected to the end of the first mobile component to achieve horizontal movement, the 8-channel pipetting component and the carrying clamp are both connected to the end of the second mobile component to achieve horizontal movement, and the 96-channel pipetting component, the 8-channel pipetting component and the carrying clamp can each move in the vertical direction, and the 96-channel pipetting component and the 8-channel pipetting component are both equipped with the pipette head.

[0007] In some possible implementations, three 8-channel pipetting assemblies capable of moving in a vertical direction are provided at the end of the second moving assembly, and the capacities of the pipette heads of the three 8-channel pipetting assemblies are different.

[0008] In some possible embodiments, it includes a consumable module and a waste bucket arranged on the frame, the consumable module includes a consumable stack and a consumable extraction mechanism, the consumable stack and the consumable extraction mechanism are both electrically connected to the monitoring component, the consumable stack is used to store the pipette head box, the pipette head box is used to store the pipette heads, the consumable extraction mechanism takes the pipette head box out of the consumable stack and lifts it to the preset position, the carrying clamp takes out the pipette head box and places it on the frame, the 96-channel pipetting assembly and the 8-channel pipetting assembly take out the pipette heads from the corresponding pipette head boxes and install the pipette heads, and the moving module docks with the waste bucket to unload the old pipette heads.

[0009] In some possible implementations, the consumables stack includes a consumables rack and a consumables motor for driving the consumables rack to rotate, and the consumables rack is provided with a plurality of installation cabins, and the installation cabins are used to place the pipette head boxes;

[0010] The consumables extraction mechanism includes a driving member, a clamp connected to the end of the driving member, and a lifting mechanism. The driving member can drive the clamp to dock with the lifting mechanism and the installation cabin respectively, so that the pipette head box is transferred to the lifting mechanism, and the lifting mechanism can dock with the transport clamp.

[0011] In some possible embodiments, the rubber cutting module includes a rubber cutting head, a blue light lamp and a visual detection component. A rubber cutting head box is provided on the frame, and a plurality of rubber cutting heads are provided in the rubber cutting head box. The first movable component can be docked with the rubber cutting head box to replaceably arrange the rubber cutting head at the end of the first movable module. The rubber cutting head and the visual detection component are both arranged at the end of the first movable component to realize horizontal movement, and the rubber cutting head can realize vertical movement; the blue light lamp is provided on the frame and can illuminate the storage module, the visual detection component is used to take pictures of the storage module, and the rubber cutting head is docked with the storage module for rubber cutting.

[0012] In some possible embodiments, the frame is provided with a substrate, the moving module is provided above the substrate, a support frame, a support plate and a third moving component are provided on the substrate, the support frame is slidably connected to the substrate and connected to the output end of the third moving component, the support plate is provided above the support frame, and the third moving component is electrically connected to the monitoring component; the moving module includes a liquid pipetting head box, and the glue cutting module includes a glue cutting head box;

[0013] A portion of the pipetting head box, the rubber cutting head box, the enzyme label concentration detection module, the electrophoresis module, the filtration purification module, the storage module, the centrifugation module, the constant temperature oscillation module, the sealing module and the PCR instrument are arranged on the substrate, and the other portions are arranged on the support plate.

[0014] In some possible embodiments, the third moving component includes a driving motor and two belt drive components, the belt drive component includes a driving wheel, a driven wheel and a belt, the belt is wound around the driving wheel and the driven wheel, the driving motor is connected to the driving wheel, the two driving wheels are connected by a rotating shaft, the support frame and the support plate form a gate shape, and the two ends of the support frame are correspondingly connected to the belts of the two belt drive components.

[0015] In some possible implementations, a protective cover is further provided on the frame, and the frame is further provided with:

[0016] a fan filter unit, the air outlet of which faces the interior of the protective cover; and / or

[0017] The UV lamp is irradiated toward the inside of the protective cover.

[0018] A gene splicing and construction method using the high-throughput gene splicing and construction workstation as described above, comprising:

[0019] Enter the preset parameters in the monitoring component to perform the following steps;

[0020] The mobile module is docked with the storage module to configure the primer;

[0021] The mobile module is docked with the storage module to configure the gene splicing reaction solution;

[0022] The mobile module is docked with the sealing module and the PCR instrument respectively to run the splicing reaction;

[0023] The mobile module is docked with the storage module to configure a gene product amplification reaction solution;

[0024] The mobile module is docked with the sealing module and the PCR instrument respectively to run the amplification reaction;

[0025] The mobile module is docked with the electrophoresis module and the storage module respectively to perform agarose gel electrophoresis;

[0026] The gel cutting module is docked with the storage module to cut the target gene band;

[0027] The mobile module is docked with the storage module, the constant temperature oscillation module, the suction filtration purification module and the centrifugal module respectively to purify the electrophoresis PCR product;

[0028] The mobile module is docked with the enzyme label concentration detection module to perform enzyme label quantification and make the concentration uniform;

[0029] The mobile module is docked with the storage module to configure the enzyme-linked reaction solution;

[0030] The mobile module is docked with the sealing module and the PCR instrument respectively to run the enzyme-linked reaction.

[0031] Beneficial effects of the present invention:

[0032] The present invention provides an automated workstation for high-throughput gene splicing and a gene splicing and construction method, which realizes the full process, automation and integrated operation of gene splicing and construction, significantly improves the experimental efficiency, and reduces manual intervention and operation steps. The above-mentioned mobile module, enzyme concentration detection module, electrophoresis module, suction filtration purification module, storage module capable of storing solvent, centrifugal module, constant temperature oscillation module, film sealing module, PCR instrument, and gel cutting module can be freely matched to meet the needs of different experimental scenarios, and improve the flexibility, applicability and scalability of the workstation. The modules work together to achieve seamless connection from gene splitting primers to plasmid construction containing target genes, thereby improving overall work efficiency and experimental success rate.

[0033] The gene splicing and construction method provided by the present invention realizes the full process, automation and integrated operation from gene splitting primers to plasmid construction containing the target gene by adopting the above-mentioned high-throughput gene splicing automated workstation, which significantly improves the experimental efficiency and reduces manual intervention and operation steps. In addition, only some of the steps can be used to perform the corresponding work, which improves the flexibility, applicability and scalability of the workstation. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 is a schematic diagram of an automated workstation for high-throughput gene splicing provided by a specific embodiment of the present invention;

[0035] Figure 2 is a structural layout diagram on a substrate provided by a specific embodiment of the present invention;

[0036] Figure 3 is a schematic diagram of a third moving assembly and a support frame provided in a specific embodiment of the present invention;

[0037] Figure 4 is a schematic diagram of a consumables module provided in a specific embodiment of the present invention;

[0038] Figure 5 It is a flow chart of the gene splicing and construction method provided by the specific embodiment of the present invention.

[0039] In the figure:

[0040] 1. frame; 11. base plate; 12. support frame; 13. support plate; 14. third moving assembly; 141. belt transmission assembly; 1411. driving wheel; 1412. driven wheel; 1413. belt; 142. guide assembly; 1421. guide rail; 1422. slider; 143. driving motor; 15. fan filter unit;

[0041] 2. Mobile module; 21. First mobile assembly; 22. Second mobile assembly; 23. 96-channel pipetting assembly; 24. 8-channel pipetting assembly; 25. Carrying gripper; 26. Pipette head; 27. Pipette head box;

[0042] 3. ELISA concentration detection module; 31, 384 ELISA plate;

[0043] 4. Electrophoresis module; 41. Peristaltic pump; 42. Electrophoresis instrument;

[0044] 5. Filtration purification module; 51. Filtration plate;

[0045] 6. Storage module; 61. Liquid storage tank; 62. PCR plate; 63. Deep well plate; 64. Waste liquid tank;

[0046] 7. Centrifugal module; 71. Centrifuge; 72. First centrifugal plate; 73. Second centrifugal plate;

[0047] 8. Constant temperature oscillation module;

[0048] 9. Film sealing module;

[0049] 10. PCR instrument;

[0050] 20. Glue cutting module; 201. Glue cutting head; 202. Visual inspection component; 203. Glue cutting head box;

[0051] 30. Consumables module; 310. Consumables stack; 3101. Consumables rack; 3102. Installation cabin; 3103. Consumables motor; 320. Consumables extraction mechanism; 3201. Driving member; 32011. First driving assembly; 32012. Second driving assembly; 3202. Clamping jaw; 3203. Lifting mechanism;

[0052] 40. Panoramic camera;

[0053] 50. Waste bin. DETAILED DESCRIPTION

[0054] In order to make the technical problems solved by the present invention, the technical solutions adopted and the technical effects achieved clearer, the technical solutions of the embodiments of the present invention will be further described in detail below in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention.

[0055] In the description of the present invention, unless otherwise clearly specified and limited, the terms "connected", "connected", and "fixed" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0056] In the present invention, unless otherwise clearly specified and limited, a first feature being "above" or "below" a second feature may include that the first and second features are in direct contact, or may include that the first and second features are not in direct contact but are in contact through another feature between them. Moreover, a first feature being "above", "above" and "above" a second feature includes that the first feature is directly above and obliquely above the second feature, or simply indicates that the first feature is higher in level than the second feature. A first feature being "below", "below" and "below" a second feature includes that the first feature is directly below and obliquely below the second feature, or simply indicates that the first feature is lower in level than the second feature.

[0057] like Figure 1-Figure 4As shown, this embodiment provides a high-throughput gene splicing and construction workstation, including a frame 1 and a mobile module 2, an enzyme label concentration detection module 3, an electrophoresis module 4, a suction filtration and purification module 5, a storage module 6 capable of storing solvents, a centrifugal module 7, a constant temperature oscillation module 8, a sealing module 9, a PCR instrument 10, a gel cutting module 20 and a monitoring component arranged on the frame 1. Among them, the monitoring component is electrically connected to the mobile module 2, the enzyme label concentration detection module 3, the electrophoresis module 4, the suction filtration and purification module 5, the storage module 6 capable of storing preset solvents, the centrifugal module 7, the constant temperature oscillation module 8, the sealing module 9, the PCR instrument 10 and the gel cutting module 20, respectively, so that the mobile module 2 can be docked with the enzyme label concentration detection module 3, the electrophoresis module 4, the suction filtration and purification module 5, the storage module 6 capable of storing solvents, the centrifugal module 7, the constant temperature oscillation module 8, the sealing module 9 and the PCR instrument 10, and the gel cutting module 20 can be docked with the storage module 6. Among them, the mobile module 2 is equipped with a liquid pipette head 26 to take and put the solvent. The monitoring component configures primers, configures gene splicing reaction solution, runs the splicing reaction, configures gene product amplification reaction solution, runs the amplification reaction, performs agarose gel electrophoresis, cuts target gene bands, purifies electrophoretic PCR products, performs enzyme labeling quantification and concentration uniformity, configures enzyme-linked reaction solution, and runs the enzyme-linked reaction according to preset information.

[0058] The full process, automation and integrated operation of gene splicing and construction are realized, which significantly improves the experimental efficiency and reduces manual intervention and operation steps. The above-mentioned mobile module 2, enzyme concentration detection module 3, electrophoresis module 4, suction filtration purification module 5, storage module 6 capable of storing solvent, centrifugal module 7, constant temperature oscillation module 8, sealing module 9, PCR instrument 10, and gel cutting module 20 can be freely matched to meet the needs of different experimental scenarios, such as using only part of the structure to perform part of the work. In the first embodiment, only the mobile module 2 and the storage module 6 are used to configure the primers. In the second embodiment, only the mobile module 2, the storage module 6 and the electrophoresis module 4 are used to perform electrophoresis. In the third embodiment, the mobile module 2, the storage module 6, the sealing module 9, the PCR instrument 10 and the electrophoresis module 4 are used to perform electrophoresis. The difference between the second embodiment and the third embodiment is that the solvents stored before using the workstation are different. The solvent stored in the second embodiment includes configured primers, etc., while the solvent stored in the third embodiment is unconfigured primers, etc., and the primers need to be configured. The specific settings are based on the needs and will not be repeated here, which improves the flexibility, applicability and scalability of the workstation. The modules work together to achieve a seamless connection from gene splitting primers to plasmid construction containing the target gene, improving the overall work efficiency and experimental success rate.

[0059] The mobile module 2 includes a first mobile component 21, a second mobile component 22, a 96-channel pipetting component 23, an 8-channel pipetting component 24 and a carrying clamp 25, all of which are electrically connected to the monitoring component. The 96-channel pipetting component 23 is connected to the end of the first mobile component 21 to achieve horizontal movement, and the 8-channel pipetting component 24 and the carrying clamp 25 are connected to the end of the second mobile component 22 to achieve horizontal movement, and the 96-channel pipetting component 23, the 8-channel pipetting component 24 and the carrying clamp 25 can each move in the vertical direction, so that the 96-channel pipetting component 23, the 8-channel pipetting component 24 and the carrying clamp 25 can be independently controlled to work. The 96-channel pipetting component 23 and the 8-channel pipetting component 24 are both equipped with a pipette head 26.

[0060] By setting the first moving assembly 21 and the second moving assembly 22, the horizontal movement of the 96-channel pipetting assembly 23, the 8-channel pipetting assembly 24 and the carrying clamp 25 is achieved, so as to facilitate the connection with other structures. Specifically, the first moving assembly 21 and the second moving assembly 22 are the same, both including an x-axis linear module and a y-axis linear module. The x-axis linear module and the y-axis linear module are both prior arts and will not be described in detail. The x-axis linear module is connected to the output end of the y-axis linear module, the 96-channel pipetting assembly 23 is connected to the output end of the x-axis linear module, and the 8-channel pipetting assembly 24 and the carrying clamp 25 are connected to the output end of another x-axis linear module.

[0061] The 8-channel pipetting assembly 24 and the 96-channel pipetting assembly 23 both include a mounting frame, a power pump, and a z-axis linear module. The mounting frame is connected to the output end of the z-axis linear module. The pipette head 26 is mounted on the mounting frame. The pipette head 26 is connected to the power pump, and the power pump controls the pipette head 26 to take and place the solvent. The mounting frame of the 8-channel pipetting assembly 24 is used to install 8 pipette heads 26, and the mounting frame of the 96-channel pipette assembly 23 is used to install 96 pipette heads 26. The transport clamp 25 includes a z-axis linear module and a clamp cylinder connected to the end thereof, and the clamp cylinder is used to achieve grasping and placing.

[0062] The end of the second moving assembly 22 is provided with three 8-channel liquid transfer assemblies 24 that can each move in the vertical direction, and the capacities of the liquid pipetting tips 26 of the three 8-channel liquid transfer assemblies 24 are different, illustratively, the pipette tips are 1000ul, 50ul and 10ul, respectively. According to the required amount of solvent, the corresponding 8-channel liquid transfer assembly 24 is selected to further improve flexibility and applicability.

[0063] The mobile module 2 also includes a pipette head box 27, which is used to store the pipette head 26. The pipette head 26 can be installed on the mobile module 2. When used for taking in and putting in different solvents, a new pipette head 26 needs to be replaced to prevent contamination caused by cross-use between different solutions.

[0064] The gel cutting module 20 includes a gel cutting head 201, a blue light lamp and a visual detection component 202. A gel cutting head box 203 is provided on the frame 1. A plurality of gel cutting heads 201 are provided in the gel cutting head box 203. The first moving component 21 can dock with the gel cutting head box 203 so that the gel cutting head 201 can be replaceably arranged at the end of the first moving component 21. The gel cutting head 201 and the visual detection component 202 are both arranged at the end of the first moving component 21 to achieve horizontal movement, and the gel cutting head 201 can achieve vertical movement. The blue light lamp is provided on the frame 1 and can illuminate the storage module 6. The visual detection component 202 is used to take pictures of the storage module 6. The gel cutting head 201 is docked with the storage module 6 for gel cutting. The blue light lamp is used for irradiation and the visual detection component 202 takes pictures to automatically identify the target DNA band, so that the gel cutting head 201 can be used for accurate gel cutting according to the identification result, thereby improving the accuracy. The cutting head 201 is installed on the first moving component 21 to facilitate docking with the storage module 6, thereby reducing the number of moving components used, simplifying the structure and reducing costs. Specifically, a z-axis linear module is also installed between the cutting head 201 and the first moving component 21, so that when cutting is required, the cutting head 201 and the 96-channel pipetting component 23 are independently controlled to perform corresponding work.

[0065] The glue cutting module 20 also includes a glue cutting head box 203, which is used to store the glue cutting head 201. The glue cutting head box 203 is used to store the glue cutting head 201, and the glue cutting head 201 can be installed on the glue cutting module 20. When used for glue cutting of different products, a new glue cutting head 201 needs to be replaced to prevent cross-use between different products and cause contamination. Exemplarily, the 8-channel pipetting assembly 24 is the same as the 96-channel pipetting assembly 23, and the 96-channel pipetting assembly 23 is used as an example for exemplary description. The 96-channel pipetting assembly 23 also includes a connecting tube and a micro pump, and the liquid suction head 26 is connected to the connecting tube, and the other end of the connecting tube is connected to the micro pump. The micro pump absorbs or releases the solvent through the liquid suction head 26. When replacing, a card plate and a cylinder driving the card plate can be provided on the 96-channel pipetting assembly 23, and the cylinder drives the card plate to move, and the card plate contacts the liquid suction head 26, and then the connecting tube moves upward to remove the liquid suction head 26 through the card plate.

[0066] The suction filtration purification module 5 includes a suction filtration plate 51 and a recovery plate, the centrifugal module 7 includes a centrifuge 71, a first centrifugal plate 72 and a second centrifugal plate 73, and the mobile module 2 can transport the suction filtration plate 51 to the recovery plate and the centrifuge 71. The electrophoresis module 4 includes an electrophoresis instrument 42 and an agarose gel plate for accommodating agarose gel. The enzyme-labeled concentration detection module 3 includes an enzyme-labeled instrument and a 384 enzyme-labeled plate 31, wherein the enzyme-labeled instrument is an electrically connected external device. The sealing module 9 includes a sealing instrument. The monitoring component includes a panoramic camera 40 and a control host. The above structures are all prior art and can be purchased from outside. For specific usage methods, please refer to the description in the subsequent gene splicing and construction method.

[0067] The frame 1 is provided with a substrate 11, the mobile module 2 is arranged above the substrate 11, the substrate 11 is provided with a support frame 12, a support plate 13 and a third mobile component 14, the support frame 12 is slidably connected to the substrate 11 and connected to the output end of the third mobile component 14, the support plate 13 is arranged above the support frame 12, and the third mobile component 14 is electrically connected to the monitoring component. A part of the liquid pipetting head box 27, the rubber cutting head box 203, the enzyme label concentration detection module 3, the electrophoresis module 4, the filtration purification module 5, the storage module 6, the centrifugal module 7, the constant temperature oscillation module 8, the sealing module 9 and the PCR instrument 10 are arranged on the substrate 11, and the other part is arranged on the support plate 13. According to actual use requirements, the third mobile component 14 drives the support frame 12 and the support plate 13 and the storage module 6 thereon to move, so that the storage module 6 located on the substrate 11 is exposed, which is convenient for docking with the mobile module 2, and the mobile module 2 is taken and placed on the storage module 6 at the corresponding exposed position. By dividing it into two parts and placing them on the base plate 11 and the support plate 13 respectively, the overall horizontal size of the workstation is reduced, the structure is compact, and the occupied space is reduced. The above structures are arranged on the base plate 11 and the support plate 13, and the mobile module 2 is arranged above the base plate 11, so that the mobile module 2 is conveniently connected to the above structures.

[0068] Specifically, the base plate 11 is provided with a first centrifugal plate placement area, a recovery plate placement area, a filtration plate placement area, an agarose gel plate placement area, a liquid pipetting head box placement area, and a gel cutting head box placement area; the support plate 13 is provided with a PCR plate 62 placement area, a 384 enzyme label plate placement area and a second centrifugal plate placement area.

[0069] Specifically, the third moving assembly 14 includes a driving motor 143, two belt transmission assemblies 141 and two guide assemblies 142. The belt transmission assembly 141 includes a driving wheel 1411, a driven wheel 1412 and a belt 1413. The belt 1413 is wound around the driving wheel 1411 and the driven wheel 1412. The driving motor 143 is connected to the driving wheel 1411 of the belt transmission assembly 141. The guide assembly 142 includes a guide rail 1421 and a slider 1422 slidably connected to the guide rail 1421. The guide rail 1421 is connected to the base plate 11. The slider 1422 is connected to the support frame 12. The support frame 12 and the support plate 13 form a gate shape. The two ends of the support frame 12 are correspondingly connected to the belts 1413 of the two belt transmission assemblies 141 and the sliders 1422 of the two guide assemblies 142. The third moving assembly 14 is arranged below the base plate 11. The support frame 12 passes through the base plate 11, which further makes the structure more compact in the vertical direction.

[0070] Exemplarily, the base plate 11 is divided into 44 working positions, wherein positions T1 to T10 are all located on the support plate 13, and the remaining positions are all located on the base plate 11. The positions and corresponding functions are shown in the following table:

[0071] Table 1 Board position and function correspondence table

[0072]

[0073] The storage module 6 includes a liquid storage tank 61, a PCR plate 62, a deep well plate 63, and a waste liquid tank 64, etc. The preset solvents that can be accommodated include pure water, electrophoresis buffer, primers, LOADING BUFFER, DNA maker, enzyme-linked reagents, carriers, PCR enzymes, sols, washing solutions, reaction solutions, reaction products, and waste liquids. The storage module 6 stores the above solvents, and any solvent can be selected for reaction, which improves applicability and flexibility. The above solvents are stored in the upper plate position, and the details are referred to the subsequent gene splicing and construction method.

[0074] The high-throughput gene splicing and construction workstation includes a consumable module 30 and a waste bucket 50 arranged on a frame 1. The consumable module 30 includes a consumable stack 310 and a consumable extraction mechanism 320. Both the consumable stack 310 and the consumable extraction mechanism 320 are electrically connected to the monitoring component. The consumable stack 310 is used to store the pipette head box 27, and the pipette head box 27 is used to store the pipette head 26. The consumable extraction mechanism 320 takes the pipette head box 27 out of the consumable stack 310 and lifts it to a preset position. The transport clamp 25 takes out the pipette head box 27 and places it on the frame 1. The 96-channel pipetting assembly 23 and the 8-channel pipetting assembly 24 are taken out from the corresponding pipette head box 27 and the pipette head 26 is installed. The mobile module 2 is docked with the waste bucket 50 to unload the old pipette head 26. The consumable stack 310 stores multiple pipette head boxes 27 for large-scale replacement.

[0075] Specifically, the substrate 11 is provided with an opening and an avoidance opening, the consumable stack 310, the consumable extraction mechanism 320 and the waste bucket 50 are all arranged on the lower side of the substrate 11, and the waste bucket 50 is passed through the avoidance opening. When the mobile module 2 is docked with the waste bucket 50, it faces the barrel opening of the waste bucket 50, and the consumable extraction mechanism 320 transfers the pipette head box 27 to the opening, and the carrying clamp 25 takes away the pipette head box 27 at the opening, that is, the opening is the docking point of the consumable module 30.

[0076] The consumable stack 310 comprises a consumable rack 3101 and a consumable motor 3103, wherein a plurality of mounting chambers 3102 are provided on the consumable rack 3101, and the mounting chambers 3102 are used to place the liquid head box 27, and the consumable motor 3103 drives the consumable rack 3101 to rotate so that the corresponding mounting chamber 3102 rotates to a preset position to dock with the consumable extraction mechanism 320. The consumable extraction mechanism 320 comprises a driving member 3201, a clamping jaw 3202 connected to its end, and a lifting mechanism 3203. The driving member 3201 drives the clamping jaw 3202 to a preset position to clamp the liquid head box 27 of the corresponding mounting chamber 3102. The lifting mechanism 3203 can dock with the carrying clamping jaw 25 and the clamping jaw 3202 respectively to realize the transfer of the liquid head box 27. The driving member 3201 may comprise a first driving assembly 32011 and a second driving assembly 32012, which respectively realize that the clamping jaw 3202 moves in the horizontal and vertical directions. Both are conventional linear drive structures such as a combination of a cylinder or a motor and a lead screw nut or a combination of a motor and a chain drive assembly. For example, the first drive assembly 32011 is a combination of a first motor and a lead screw nut, the nut is connected to a fixed plate, the clamp cover is fixed on the fixed plate, the first motor is fixed at the output end of the second drive assembly 32012, the second drive assembly 32012 is a second motor and a chain drive assembly, the chain drive assembly includes a transmission chain for transmission, the first motor and the lead screw are fixed on the transmission chain, the second motor drives the transmission chain forward and reversely to make the transmission chain reciprocate, thereby driving the first drive assembly 32011 to reciprocate in the vertical direction.

[0077] Specifically, the clamp 3202 places the liquid head box 27 on the lifting mechanism 3203, and the lifting mechanism 3203 lifts the liquid head box 27 to the opening to facilitate the carrying clamp 25 to grab. The lifting mechanism 3203 includes a second linear drive member 3201 and a lifting plate installed at the end thereof. The first linear drive member 3201 and the second linear drive member 3201 can be a cylinder or a combination of a motor and a lead screw nut, which is a prior art and will not be repeated.

[0078] The high-throughput gene splicing and construction workstation also includes a protective cover arranged on the frame 1 to form a clean room inside. The frame 1 is also provided with: a fan filter unit and a UV lamp. The full name of the fan filter unit 15 is Fan Filter Unit, abbreviated as FFU in English. It is an air filter system with a built-in fan. The air outlet of the fan filter unit 15 faces the inside of the protective cover. The fan filter unit 15 can provide static pressure to keep the clean room positive pressure relative to the outside world, prevent external particles from penetrating into the clean area, ensure sealing and safety, and ensure that the environment inside the equipment is in a pollution-free state during the operation of the automated process. The UV lamp is an ultraviolet lamp, which irradiates toward the inside of the protective cover to achieve the purpose of disinfection, sterilization and cleaning.

[0079] like Figure 5As shown, this embodiment also provides a gene splicing and construction method, using the above-mentioned high-throughput gene splicing and construction workstation, including the following steps:

[0080] Before using the high-throughput gene splicing and construction workstation, manually place the consumables and reagents according to the positions in Tables 2 and 3 below to ensure that the reagents and consumables are placed correctly.

[0081] Table 2 Consumables placement table

[0082]

[0083]

[0084] Table 3 Reagent placement table

[0085]

[0086]

[0087] S1. According to the needs, input the preset parameters in the monitoring component to perform the following steps. Specifically, manually import the parameter table required for the monitoring component: including the gene parameter input table and the transfer vector parameter table, and start the system. The system will automatically complete the following operations. Among them, the gene parameter input table: contains DNA content and sequence length information, which is used for the target band identification link after electrophoresis gel running in the process, and the concentration uniformity link after product recovery. Transfer vector parameter table: contains the vector information required for each construction, which is used for the gene fragment enzyme connection to the target vector link in the process.

[0088] S2, the mobile module 2 is docked with the storage module 6 to configure the primers; specifically, the mobile module 2 includes a 96-channel pipetting assembly 23 and a transporting clamp 25. The 96-channel mobile module 2 transfers the pure water from the T20 liquid storage plate to the T17 deep well plate 63 containing the first primer and the T18 deep well plate 63 containing the tail primer to dilute the primers, and then mixes the diluted first primer in the T17 deep well plate 63 and the diluted tail primer in the T18 deep well plate 63 in equal proportions into the T28 PCR plate 62. The transporting clamp 25 transports the T28 PCR plate 62 containing the first and tail primer mixture to the T37 well plate sealing instrument for sealing, and then transfers it to the T4 normal temperature PCR plate 62 to place the plate for standby. In this process, the 96-channel pipetting assembly 23 and the transporting clamp 25 automatically discard the discarded liquid pipetting head 26 and the liquid pipetting head box 27 into the T36 discard bucket 50. The T41 consumable module 30 automatically lifts the pipette head box 27 at the docking point, and the transport clamp 25 automatically takes away the pipette head box 27 and transports it to the designated position (T29-T34 / T45-46).

[0089] S3, the mobile module 2 is docked with the storage module 6 to configure the gene splicing reaction solution; the transport gripper 25 transports the PCR plate 62 from T7 to the T28 plate position, which is the 4°C reaction solution configuration work area, and the 8-channel pipetting component 24 of the mobile module 2 distributes the PCR enzyme from the T26 deep-well plate 63 to the T27 PCR plate 62 (the first column) in the form of 1 suction and multiple release, and then evenly distributes the PCR enzyme in the first column to the T28 PCR plate 62. The 96-channel pipetting component 23 transfers the pure water from the T20 reservoir 61 to the T25 PCR plate 62 in the 4°C reaction solution configuration work area, and then transfers the template primer from the T19 deep-well plate 63 to the T25 PCR plate 62 for mixing and dilution, and then transfers the template primer from the T25 PCR plate 62 to the T28 PCR plate 62 for mixing. In this process, the 8-channel pipetting assembly 24 and the transporting clamp 25 automatically discard the discarded pipette tips 26 and the pipette tip box 27 into the T36 waste bucket 50. The T41 consumable module 30 automatically lifts the pipette tip box 27 at the docking point, and the transporting clamp 25 automatically transports the pipette tip box 27 to the designated position (T29-T34 / T45-46).

[0090] S4, the mobile module 2 is docked with the sealing module 9 and the PCR instrument 10 respectively to run the splicing reaction; the PCR instrument 10 of T39 automatically opens, and the carrying gripper 25 carries the PCR plate 62 of T28 to the sealing instrument of T37 for sealing, and then carries it to the PCR instrument 10 of T39. The PCR instrument 10 automatically closes and automatically starts running the splicing thermal cycle program. After the reaction program is completed, the PCR instrument 10 automatically opens.

[0091] S5, the mobile module 2 is docked with the storage module 6 to configure the gene product amplification reaction solution; the 8-channel pipetting component 24 evenly distributes the PCR enzymes from the 2nd to the 4th columns of T27 to the PCR plate 62 of T28. The transporting clamp 25 transfers the PCR plate 62 of the spare first and tail primer mixture of T4 to the T10 plate position (closer to the T28 plate position, which can save pipetting time). The 8-channel pipetting component 24 transfers the required first and tail primer mixture from the PCR plate 62 of T10 to the PCR plate 62 of T28. After the PCR reaction program in step S4 is completed, the transporting clamp 25 transports the PCR plate 62 in the PCR instrument 10 to the T8 normal temperature PCR plate 62 placement plate position, and the 8-channel pipetting component 24 transfers the spliced ​​products in the PCR plate 62 of T8 to the PCR plate 62 of T28 as required for mixing by blowing. In this process, the 8-channel pipetting assembly 24 and the transporting clamp 25 automatically discard the discarded pipette tips 26, the pipette tip box 27 and the T10 PCR plate 62 into the T36 waste bucket 50. The T41 consumable module 30 automatically lifts the pipette tip box 27 at the docking point, and the transporting clamp 25 automatically transports the pipette tip box 27 to the designated position (T29-T34 / T45-46).

[0092] S6, the mobile module 2 is docked with the sealing module 9 and the PCR instrument 10 respectively to run the amplification reaction; the PCR instrument 10 of T39 automatically opens, and the carrying gripper 25 carries the PCR plate 62 of T28 to the sealing instrument of T37 for sealing, and then carries it to the PCR instrument 10. The PCR instrument 10 automatically closes and automatically starts to run the amplification thermal cycle program. After the reaction program is completed, the PCR instrument 10 automatically opens.

[0093] S7, the mobile module 2 is docked with the electrophoresis module 4 and the storage module 6 respectively to perform agarose gel electrophoresis; the transport gripper 25 transports the PCR plate 62 in the PCR instrument 10 to the T10 room temperature PCR plate 62 placement plate position. The large liquid storage tank 61 begins to pump the electrophoresis buffer into the 96-channel agarose gel electrophoresis tank of T11 through the peristaltic pump 41. After the 8-channel pipetting component 24 transfers the loading buffer from the PCR plate 62 (column 5) of T27 to the PCR product of the PCR plate 62 of T10, the PCR product is slowly pumped into the 96-channel agarose gel electrophoresis tank of T11, and then the DNA Marker is pumped from the PCR plate 62 of T27 (hole A12) into the designated gel hole in the form of 1 suction and multiple release. The electrophoresis instrument 42 performs electrophoresis, and after the electrophoresis is completed, the electrophoresis is automatically stopped, and the electrophoresis buffer in the 96-channel agarose gel electrophoresis tank is pumped out into the large liquid storage tank 61.

[0094] S8, the gel cutting module 20 is docked with the storage module 6 to cut the target gene band; the gel cutting blue light is automatically turned on, and the industrial camera of the visual detection component 202 automatically moves to the photo taking position to collect images and automatically identifies the target DNA band (the position of the band identification can also be manually adjusted). According to the identification result, the gel cutting module 20 uses the gel cutting head 201 to accurately cut the gel (insert into the gel and then suck out the gel), and then transfers one by one to the corresponding empty space of the T21 deep well plate 63.

[0095] S9, the mobile module 2 is docked with the storage module 6, the constant temperature oscillation module 8, the filtration purification module 5 and the centrifugal module 7 respectively to purify the electrophoresis PCR product; the 8-channel pipetting assembly 24 transfers the required amount of sol liquid from the four liquid adding tanks of T24 (the first column) to the T21 deep well plate 63. The transport clamp 25 transports the T21 deep well plate 63 to the T38 constant temperature oscillator for 50 degrees constant temperature oscillation to dissolve the agarose gel block in the hole. After the gel is melted, the system automatically moves the deep well plate 63 back to the T21 plate position. The 8-channel pipetting assembly 24 transfers all the liquid in the T21 deep well plate 63 to the T42 suction plate 51 in batches. After standing for 3 minutes, the negative pressure suction filtration is automatically turned on and off. Next, the 8-channel pipetting assembly 24 transfers the required amount of washing liquid from the four liquid adding tanks of T24 (the second column) to the T42 suction plate 51, and the negative pressure suction filtration is automatically turned on and off. The 8-channel pipetting assembly 24 transfers the required amount of washing liquid from the T24 four-channel liquid adding tank (column 3) to the T42 filter plate 51 again, and the negative pressure filtration is automatically turned on and off again. The transport clamp 25 transports the filter plate 51 to the T9 recovery plate (for receiving waste liquid), and then places it in the centrifuge 71 for centrifugation for 10 minutes. After the centrifugation is completed, the recovery plate is manually removed and the filter plate 51 is placed on the T35 recovery plate (to receive the purified product). The 96-channel pipetting assembly 23 synchronously transfers the required amount of pure water from the T20 liquid storage plate to the T35 filter plate 51. After standing for 5 minutes, it is placed in the centrifuge 71 for centrifugation for 10 minutes to elute the purified product.

[0096] S10, mobile module 2 is docked with enzyme label concentration detection module 3 to carry out enzyme label quantification and make concentration uniform; remove the suction filter plate 51, and put the recovery plate back to the T9 centrifugal plate placement plate position. The 8-channel pipetting assembly 24 transfers the purified PCR product to the 384 enzyme label plate 31 of T2 in sequence, and adds pure water as a control group. At this time, the microplate reader will open the warehouse, and the system will alarm and prompt to manually transfer the 384 enzyme label plate 31 to the microplate reader (connection system) for concentration detection, and remind to put the 384 enzyme label plate 31 back to the T2 placement plate position. The system automatically calculates the amount of water needed to add for uniform sample concentration according to the information of the "gene parameter input table", and directly issues a command to control the 8-channel pipetting assembly 24 to add pure water to the corresponding sample as needed from the T20 liquid storage plate. Specifically, an 8-channel pipetting assembly 24 of corresponding capacity can be selected to take and put pure water.

[0097] S11, the mobile module 2 is docked with the storage module 6 to configure the enzyme-linked reaction solution; the transport gripper 25 transports the consumables of the PCR plate 62 of T6 to the plate position of the 4°C reaction solution configuration work area of ​​T28, and the system transfers the required amount of linearized vector from the PCR plate 62 of T27 (columns 7-12) to the corresponding well position of the PCR plate 62 of T28 according to the information of the "transfer carrier parameter table", and then transfers the PCR product after the concentration of the 384 ELISA plate 31 of T2 to the corresponding well position of the PCR plate 62 of T28. Finally, the enzyme-linked reagent is transferred from the PCR plate 62 of T27 (column 6) to the PCR plate 62 of T28 according to the required amount and mixed by reverse pipetting.

[0098] S12, the mobile module 2 is docked with the sealing module 9 and the PCR instrument 10 respectively to run the enzyme-linked reaction. The PCR instrument 10 of T39 automatically opens, and the carrying clamp 25 carries the PCR plate 62 of T28 to the sealing instrument of T37 for sealing, and then carries it to the PCR instrument 10. The PCR instrument 10 automatically closes and automatically starts to run the enzyme-linked reaction program. After the reaction program is completed, the PCR instrument 10 automatically opens.

[0099] Through the above steps, the whole process, automation and integrated operation from gene splitting primer to plasmid construction containing target gene is realized, which significantly improves the experimental efficiency and reduces manual intervention and operation steps. During use, only part of the structure and only the work of opening the corresponding steps can be used to perform part of the work. In the first embodiment, the mobile module 2 and the storage module 6 are used, and steps S1 and S2 are used to configure the primers. In the second embodiment, the mobile module 2, the storage module 6 and the electrophoresis module 4 are used, and steps S7 and S8 are used to perform electrophoresis. In the third embodiment, the mobile module 2, the storage module 6, the sealing module 9, the PCR instrument 10 and the electrophoresis module 4 are used, and steps S1 to S8 are used to perform electrophoresis. The difference between the second embodiment and the third embodiment is that the solvents stored before using the workstation are different. The solvent stored in the second embodiment includes the primers that have been configured, etc., while the solvent stored in the third embodiment is the primers that are not configured, etc., and the primers need to be configured. It is specifically set according to the needs, and no further description is given, which improves the flexibility, applicability and scalability of the workstation.

[0100] Furthermore, the above high-throughput gene splicing and construction process is completed and also includes the following step S12, the system reminds, such as setting an alarm to realize an alarm, to remind the PCR plate 62 in the microplate reader to be manually transferred to the automated competent cell transformation workstation for transformation, and to clean up all remaining consumables and reagents in the equipment.

[0101] In the above process, the 96-channel pipetting assembly 23, the 8-channel pipetting assembly 24 and the transport clamp 25 will automatically discard the discarded pipette tips 26, the pipette tip box 27 and the PCR plate 62 into the T36 waste bucket 50. The T41 consumable module 30 will automatically lift the pipette tip box 27 at the docking point, and the transport clamp 25 will automatically move the pipette tip box 27 to the designated position (T29-T34 / T45-46). Before using the reagents in the T27 plate position, the system will automatically move the upper cover (to prevent the generation of condensate) to the T22 plate position, and the system will automatically move the cover back after use.

[0102] The above process can be run automatically and continuously from beginning to end, or it can be set to run in single steps. Change the reaction conditions or adjust the process steps according to the actual experimental needs.

[0103] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the embodiments here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. A high-throughput gene splicing and construction workstation, characterized in that: The invention comprises a frame (1), on which are provided: a mobile module (2), an enzyme-labeled concentration detection module (3) capable of docking with the mobile module (2), an electrophoresis module (4), a filtration purification module (5), a storage module (6) capable of storing a preset solvent, a centrifugation module (7), a constant temperature oscillation module (8), a film sealing module (9) and a PCR instrument (10); the frame (1) is also provided with a gel cutting module (20) and a monitoring component, the gel cutting module (20) being capable of docking with the storage module (6), the monitoring component being capable of being electrically connected to the above structures, and configuring primers, configuring gene splicing reaction liquid, running the splicing reaction, configuring gene product amplification reaction liquid, running the amplification reaction, agarose gel electrophoresis, cutting target gene bands, electrophoresis PCR product purification, enzyme-labeled quantification and concentration uniformity, configuring enzyme-linked reaction liquid and running the enzyme-linked reaction; the mobile module (2) is provided with a liquid pipette head (26) for taking and placing the solvent.

2. The high-throughput gene splicing and construction workstation according to claim 1, characterized in that: The moving module (2) comprises a first moving component (21), a second moving component (22), a 96-channel pipetting component (23), an 8-channel pipetting component (24) and a carrying clamp (25), all of which are electrically connected to the monitoring component. The 96-channel pipetting component (23) is connected to the end of the first moving component (21) to achieve horizontal movement, the 8-channel pipetting component (24) and the carrying clamp (25) are both connected to the end of the second moving component (22) to achieve horizontal movement, and the 96-channel pipetting component (23), the 8-channel pipetting component (24) and the carrying clamp (25) are each capable of moving in a vertical direction, and the 96-channel pipetting component (23) and the 8-channel pipetting component (24) are both equipped with the pipette head (26).

3. The high-throughput gene splicing and construction workstation according to claim 2, characterized in that: The end of the second moving component (22) is provided with three 8-channel pipetting components (24) which can each move in a vertical direction, and the capacities of the pipetting heads (26) of the three 8-channel pipetting components (24) are different.

4. The high-throughput gene splicing and construction workstation according to claim 2, characterized in that: The consumables module (30) and the waste bucket (50) are arranged on a frame (1), wherein the consumables module (30) comprises a consumables stack (310) and a consumables extraction mechanism (320), the consumables stack (310) and the consumables extraction mechanism (320) are both electrically connected to the monitoring component, the consumables stack (310) is used to store a pipette head box (27), the pipette head box (27) is used to store a pipette head (26), and the consumables extraction mechanism (320) extracts the pipette head (26) from the pipette head. The head box (27) is taken out from the consumables stack (310) and lifted to a preset position, the carrying clamp (25) takes out the pipette head box (27) and places it on the frame (1), the 96-channel pipetting assembly (23) and the 8-channel pipetting assembly (24) are taken out from the corresponding pipette head box (27) and the pipette head (26) is installed, and the moving module (2) is docked with the waste bucket (50) to unload the old pipette head (26).

5. The high-throughput gene splicing and construction workstation according to claim 4, characterized in that: The consumables stack (310) comprises a consumables rack (3101) and a consumables motor (3103) for driving the consumables rack (3101) to rotate, and a plurality of installation chambers (3102) are provided on the consumables rack (3101), and the installation chambers (3102) are used to place the liquid pipetting head boxes (27); The consumable material extraction mechanism (320) includes a driving member (3201), a clamp (3202) connected to the end of the driving member (3201) and a lifting mechanism (3203). The driving member (3201) can drive the clamp (3202) to dock with the lifting mechanism (3203) and the installation cabin (3102) respectively, so that the liquid suction head box (27) is transferred to the lifting mechanism (3203), and the lifting mechanism (3203) can dock with the transport clamp (25).

6. The high-throughput gene splicing and construction workstation according to claim 2, characterized in that: The rubber cutting module (20) comprises a rubber cutting head (201), a blue light lamp and a visual detection component (202); a rubber cutting head box (203) is provided on the frame (1); a plurality of rubber cutting heads (201) are provided in the rubber cutting head box (203); the first movable component (21) can be docked with the rubber cutting head box (203) so that the rubber cutting head (201) can be replaceably arranged at the end of the first movable component (21); the rubber cutting head (201) and the visual detection component (202) are both arranged at the end of the first movable component (21) to achieve horizontal movement, and the rubber cutting head (201) can achieve vertical movement; the blue light lamp is arranged on the frame (1) and can illuminate the storage module (6); the visual detection component (202) is used to take pictures of the storage module (6); the rubber cutting head (201) is docked with the storage module (6) for rubber cutting.

7. The high-throughput gene splicing and construction workstation according to claim 1, characterized in that: The frame (1) is provided with a base plate (11), the moving module (2) is arranged above the base plate (11), the base plate (11) is provided with a support frame (12), a support plate (13) and a third moving component (14), the support frame (12) is slidably connected to the base plate (11) and connected to the output end of the third moving component (14), the support plate (13) is arranged above the support frame (12), and the third moving component (14) is electrically connected to the monitoring component; the moving module (2) includes a liquid suction head box (27), and the glue cutting module (20) includes a glue cutting head box (203); Part of the pipetting head box (27), the rubber cutting head box (203), the enzyme label concentration detection module (3), the electrophoresis module (4), the suction filtration purification module (5), the storage module (6), the centrifugation module (7), the constant temperature oscillation module (8), the sealing module (9) and the PCR instrument (10) are arranged on the base plate (11), and the other parts are arranged on the support plate (13).

8. The high-throughput gene splicing and construction workstation according to claim 7, characterized in that: The third moving component (14) comprises a driving motor (143) and two belt transmission components (141); the belt transmission component (141) comprises a driving wheel (1411), a driven wheel (1412) and a belt (1413); the belt (1413) is wound around the driving wheel (1411) and the driven wheel (1412); the driving motor (143) is connected to the driving wheel (1411); the two driving wheels (1411) are connected via a rotating shaft; the support frame (12) and the support plate (13) form a gate shape; the two ends of the support frame (12) are correspondingly connected to the belts (1413) of the two belt transmission components (141).

9. The high-throughput gene splicing and construction workstation according to claim 1, characterized in that: It also includes a protective cover arranged on the frame (1), and the frame (1) is also provided with: a fan filter unit (15), the air outlet of which faces the interior of the protective cover; and / or The UV lamp is irradiated toward the inside of the protective cover.

10. A gene splicing and construction method using the high-throughput gene splicing and construction workstation according to any one of claims 1 to 9, characterized in that: include: Enter the preset parameters in the monitoring component to perform the following steps; The mobile module (2) is docked with the storage module (6) to configure the primers; The mobile module (2) is docked with the storage module (6) to configure a gene splicing reaction solution; The mobile module (2) is docked with the sealing module (9) and the PCR instrument (10) respectively to run the splicing reaction; The mobile module (2) is docked with the storage module (6) to configure a gene product amplification reaction solution; The mobile module (2) is docked with the sealing module (9) and the PCR instrument (10) respectively to run an amplification reaction; The mobile module (2) is docked with the electrophoresis module (4) and the storage module (6) respectively to perform agarose gel electrophoresis; The gel cutting module (20) is docked with the storage module (6) to cut out the target gene band; The mobile module (2) is docked with the storage module (6), the constant temperature oscillation module (8), the suction filtration purification module (5) and the centrifugal module (7) respectively to purify the electrophoresis PCR product; The mobile module (2) is docked with the enzyme label concentration detection module (3) to perform enzyme label quantification and make the concentration uniform; The mobile module (2) is docked with the storage module (6) to configure the enzyme-linked reaction solution; The mobile module (2) is docked with the sealing module (9) and the PCR instrument (10) respectively to run the enzyme-linked reaction.

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