Full-automatic sequencing library building instrument and control method thereof

Abstract

The invention discloses a full-automatic sequencing library building instrument and a control method thereof. The library building instrument comprises a cover opening assembly, the cover opening assembly synchronously abuts against a lower turning part and an upper turning part of a turning cover through a first abutting part, so that the turning cover enters a semi-open or micro-open state to balance the pressure difference, and the turning cover is completely opened by pressing the lower turning part downwards through a second abutting part. When the cover is closed, the groove-shaped structure on the surface of the turning cover is used for physically guiding the abutting piece, and accurate buckling is ensured. According to the control method, the biochemical process and the step-by-step switch cover logic depth are nested. The problems of liquid splashing and pollution in the automatic cover opening process are effectively solved, the cover closing reliability and the experiment safety are improved, and standardized automatic operation of the whole process from nucleic acid extraction to library construction is achieved.

Description

Technical Field

[0001] This application relates to the fields of biomedical devices and laboratory automation technology, and in particular to a fully automated sequencing library preparation instrument and its control method. Background Technology

[0002] In the field of gene sequencing, library construction is a crucial step that determines sequencing quality. Current nucleic acid extraction and library construction processes largely rely on manual or semi-automated equipment, which suffers from drawbacks such as cumbersome operation, susceptibility to sample cross-contamination, and aerosol contamination. With the widespread adoption of high-throughput sequencing, developing highly automated library construction equipment has become an industry trend.

[0003] However, existing automated equipment still faces significant technical bottlenecks when handling capped reagent kits. On the one hand, after undergoing heating incubation or biochemical reactions, the internal pressure of the reagent kit is often higher than that of the external environment. Traditional direct opening methods can easily cause liquid splashing, resulting in environmental pollution and threatening experimental safety. On the other hand, existing automatic capping mechanisms have limited precision, often resulting in loose caps or positional misalignments during high-frequency operation, leading to reagent evaporation and concentration inaccuracies. Therefore, designing a fully automated sequencing library preparation scheme that can effectively prevent splashing while ensuring accurate capping is a problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0004] To address the shortcomings of existing technologies, according to one aspect of this application, a fully automated sequencing library preparation instrument is provided, comprising: The frame, the pipetting module and the incubation and transfer module are installed in the frame; The incubation transport module is used to carry the reagent kit; The pipetting module includes a multi-axis motion mechanism, a pipetting module, and a cap opening assembly; A multi-axis motion mechanism is connected to the pipetting module and the cap opening assembly, used to drive the pipetting module and the cap opening assembly to move relative to the incubation and transfer module; wherein... The lid opening assembly includes an abutment for performing a press-to-open the lid; When the contact element presses down to open the lid, it contacts the lid of the reagent kit, causing the lid to first flip to a half-open or slightly open state, and then move to a fully open state.

[0005] In an optional embodiment, the cover opening assembly further includes a retaining base; an abutment extends along the downward opening direction of the retaining base; the abutment includes a first abutment portion and a second abutment portion; The first abutting part is used to press down the upper and lower flip parts of the flip cover when it is in the closed state, so that the flip cover is opened to a half-open or slightly open state. The second abutment part is used to press down the lower part of the flip cover when it is in a half-open or slightly open state, so that the flip cover is opened to the fully open state.

[0006] In an optional embodiment, the first abutting portion includes a first rod and a second rod, and the second abutting portion includes a third rod; The first lever is used to press down the downward flipping part to trigger opening, and the second lever abuts against the popping upward flipping part to maintain the half-open or slightly open state together. The third lever is used to continue pressing down the flip-up part after the cover is in a half-open or slightly open state, so as to drive the flip cover to the fully open state.

[0007] In an optional embodiment, the first abutting portion and the second abutting portion are different contact surfaces at the bottom end of the abutting member; When the first contact part performs the downward stroke of the flip cover, it can contact the downward flip part and the upward flip part, so that the flip cover can be flipped to a half-open or slightly open state; When the second contact part performs the downward stroke of the flip cover, it can contact the flip part so that the flip cover is opened to the fully open state.

[0008] In an optional embodiment, the abutment includes a third abutment portion and a fourth abutment portion with independently adjustable telescopic lengths; the multi-axis motion mechanism switches the open state of the flip cover by driving the third abutment portion and the fourth abutment portion to extend to different lengths. The third abutment extends by a first extension to abut the lower part of the flip cover, while the fourth abutment extends by a second extension to abut the upper part, so as to execute the flip cover to first flip to a half-open or slightly open state. The third abutment extends by a third elongation to abut the lower part of the flip cover, while the fourth abutment disengages from the upper part to open the flip cover to the fully open state; wherein the third elongation, the first elongation, and the second elongation decrease sequentially.

[0009] According to another aspect of this application, a control method for a fully automated sequencing library preparation instrument is provided, applied to the fully automated sequencing library preparation instrument of the foregoing embodiments. The fully automated sequencing library preparation instrument includes the following steps during the process of nucleic acid extraction or library construction: The opening component is controlled to perform a first pressing action, which simultaneously abuts against the lower and upper flip parts of the reagent kit cap, so that the cap is pressed down to a half-open or slightly open state; the opening component is controlled to perform a second pressing action, so that the cap moves from the half-open or slightly open state to the fully open state. Perform the current target action of nucleic acid extraction or library construction; Control the lid opening component to close the flip cover.

[0010] In one optional embodiment, the cover opening assembly includes a first rod, a second rod, and a third rod, wherein the length of the third rod is greater than the length of the first rod, and the length of the first rod is greater than the length of the second rod. The opening assembly is controlled to perform the first pressing action, simultaneously abutting the lower and upper flip parts of the reagent kit cap to open the cap to a half-open or slightly open state. The specific steps include: The multi-axis motion mechanism is controlled to drive the opening assembly to move to the first opening position, so that the first rod is aligned with the lower part of the reagent kit cover and the second rod is aligned with the upper part of the reagent kit cover. The opening assembly is controlled to perform a first pressing action, so that the first rod abuts against the lower part of the reagent kit cover and the second rod abuts against the upper part of the reagent kit cover, so that the cover is pressed down to open to a half-open or slightly open state. "Controlling the opening assembly to perform a second pressing action, so that the flip cover moves from a half-open or slightly open state to a fully open state, specifically includes the following steps:" The multi-axis motion mechanism is controlled to drive the opening assembly to the second opening position so that the third rod is aligned with the lower part of the reagent kit lid. The control unit performs a second pressing action so that the third rod abuts against the lower part of the reagent kit cap, thereby pressing the cap down to the fully open state.

[0011] In an optional embodiment, controlling the opening assembly to close the flip cover specifically includes the following steps: The control panel lifts the lid opening assembly at the second open position so that the bottom of the third rod is aligned with the top surface of the lid when it is closed. The multi-axis motion mechanism is controlled to drive the third rod to the closed position located on the surface of the upper flip section to close the flip cover.

[0012] In an optional embodiment, the abutment includes a third abutment portion and a fourth abutment portion whose telescopic length can be independently adjusted; Before the step of controlling the cap-opening component to perform the first pressing action, and simultaneously abutting the lower and upper flip parts of the reagent kit cap to open the cap to a half-open or slightly open state, the following steps are also included: Control the opening assembly to move and align with the flip cover, so that the third abutment is aligned with the lower part of the flip cover and the fourth abutment is aligned with the upper part of the flip cover. The opening assembly is controlled to perform the first pressing action, simultaneously abutting against the lower and upper flip parts of the reagent kit cap, so that the cap is pressed open to a half-open or slightly open state. Specific steps include: The third abutment part is controlled to extend by a first extension to abut the lower flip-up part, and the fourth abutment part is simultaneously controlled to extend by a second extension to abut the upper flip-up part, so that the flip cover is pressed down and opened to a half-open or slightly open state; wherein, the first extension is greater than the second extension. The control mechanism for the lid opening assembly performs a second pressing action, causing the lid to move from a half-open or slightly open state to a fully open state. This includes the following steps: The third abutment part is controlled to extend from the first extension amount to the third extension amount, and the fourth abutment part is controlled to disengage from the flip-up part so that the flip cover is pressed down and opened to the fully open state.

[0013] In an optional embodiment, controlling the opening assembly to close the flip cover specifically includes the following steps: The opening assembly is controlled to lift, and the third abutment is switched to the third extension so that the bottom height of the third abutment is aligned with the top height of the flip cover when it is closed. The multi-axis motion mechanism is controlled to drive the third abutment part to the closed position located on the surface of the upper flip part to close the flip cover.

[0014] Compared with the prior art, the beneficial effects of the present invention are as follows: The opening assembly enables step-by-step control of the flip-top opening (partial or slightly open and fully open), and the first contact part synchronously limits the upward and downward flip-top parts, effectively solving the problem of liquid splashing and aerosol contamination caused by internal and external pressure differences after incubation or PCR reaction of the reagent kit.

[0015] By driving the deep collaboration of various functional modules through a multi-axis motion mechanism, the entire process from nucleic acid extraction to library construction is automated, which significantly reduces the labor intensity of personnel and the risk of cross-contamination of samples, and improves the consistency of experimental products.

[0016] The above description is merely an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention and to implement it according to the contents of the specification, the preferred embodiments of the present invention are described in detail below with reference to the accompanying drawings. Specific embodiments of the present invention are given in detail below with reference to the accompanying drawings. Attached Figure Description

[0017] The accompanying drawings, which are included to provide a further understanding of the invention and form part of this application, illustrate exemplary embodiments of the invention and, together with their description, serve to explain the invention and do not constitute an undue limitation thereof. In the drawings: Figure 1 This is a schematic diagram of the overall structure of the fully automated sequencing library preparation instrument provided in the embodiments of this application; Figure 2 This is a schematic diagram of the three-dimensional structure of the outer shell of the fully automated sequencing library preparation instrument provided in the embodiments of this application; Figure 3 A three-dimensional structural schematic diagram from another perspective of the fully automated sequencing library preparation instrument provided in the embodiments of this application; Figure 4A schematic diagram of the base plate and internal component layout of the fully automated sequencing library preparation instrument provided in the embodiments of this application; Figure 5 This is a schematic diagram of the pipetting module and multi-axis motion mechanism provided in the embodiments of this application; Figure 6 This is a schematic diagram of the structure of the cover opening component provided in an embodiment of this application; Figure 7 This is a schematic diagram of the pipetting module provided in the embodiments of this application; Figure 8 This is a schematic diagram illustrating the opening component provided in this application performing a first pressing action to a half-open or slightly open state; Figure 9 A schematic diagram illustrating the second pressing action of the cover opening component provided in this application embodiment to the fully opened state; Figure 10 This is a schematic diagram of the structure of the incubation transmission module provided in the embodiments of this application; Figure 11 This is an exploded view of the incubation module provided in the embodiments of this application; Figure 12 This is a schematic diagram of the structure of the magnetic suction module provided in the embodiments of this application; Figure 13 This is a schematic diagram illustrating the operation of a multi-machine parallel fully automated sequencing library preparation instrument provided in the embodiments of this application; Figure 14 This is a schematic diagram showing the detailed structure of the reagent kit flip-top provided in an embodiment of this application; Figure 15 A flowchart illustrating the fully automated sequencing library preparation instrument control method provided in this application embodiment.

[0018] In the diagram: outer shell 1, door 11, filtration system 110, ultraviolet disinfection lamp 114, pipetting module 2, opening assembly 217, Z-axis first slider 2171, Z-axis first motor 2172, Z-axis first motor fixing plate 2173, fixing seat 2174, second rod 2175, first rod 2176, third rod 2177, pipetting module 218, incubation and transfer module 3, base plate 4, flip cover 52, lower flip part 521, upper flip part 522 and groove structure 523. Detailed Implementation

[0019] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0020] In the accompanying drawings, shapes and dimensions may be enlarged for clarity, and the same reference numerals will be used in all figures to indicate the same or similar parts.

[0021] In the following description, terms such as center, thickness, height, length, front, back, rear, left, right, top, bottom, upper, and lower are defined relative to the structure shown in the accompanying drawings. In particular, "height" corresponds to the dimension from top to bottom, "width" corresponds to the dimension from left to right, and "depth" corresponds to the dimension from front to back. These are relative concepts and may vary depending on their location and usage. Therefore, these or other orientations should not be interpreted as restrictive terms.

[0022] Terms involving attachment, connection, etc. (e.g., “connection” and “attachment”) refer to the relationship in which these structures are directly or indirectly fixed or attached to each other through an intermediate structure, as well as movable or rigid attachments or relationships, unless otherwise explicitly stated.

[0023] Example 1 According to one aspect of the embodiments of this application, a fully automated sequencing library preparation instrument is provided. In some embodiments, such as Figure 1 As shown, the fully automated sequencing library preparation instrument includes a frame, a pipetting module 2 and an incubation and transfer module 3 disposed within the frame. It should be noted that, in this application, the frame refers to an overall support frame consisting of a base plate 4 and a shell 1 fixed to the base plate 4, supporting structures, etc.

[0024] Specifically, such as Figure 10 , Figure 11 As shown, the incubation transport module 3 is used to carry the reagent kit 5. In some embodiments, the reagent kit 5 is used as a consumable and is placed on the incubation transport module.

[0025] It should be noted that the specific number and specifications of the reagent kits 5 arranged on the incubation transport module can be adjusted according to experimental throughput requirements, and should not be construed as limiting the scope of protection of this application. Optionally, the incubation transport module includes an incubation module base plate 32, which is fixed on the second slider 31 in the X direction to drive the reagent kits 5 to move in the X direction, thereby realizing the automated placement or retrieval of consumables.

[0026] Specifically, such as Figure 5As shown in the embodiment of this application, the pipetting module 2 includes a multi-axis motion mechanism, a pipetting module 218, and a capping assembly 217. The multi-axis motion mechanism is connected to the pipetting module 218 and the capping assembly 217, and is used to drive the pipetting module 218 and the capping assembly 217 to move relative to the incubation and transfer module, so as to achieve precise positioning and action execution between different workstations.

[0027] It should be noted that the specific driving method of the multi-axis motion mechanism includes, but is not limited to, lead screw drive, synchronous belt drive, or linear motor drive, as long as it can achieve the positioning of the pipetting module 218 and the cap opening assembly 217 in three-dimensional space. Specifically: the multi-axis motion mechanism drives the relevant components to move in the X direction through the X-axis motor 23; the multi-axis motion mechanism drives the relevant components to move in the Y direction through the Y-axis motor 211; the cap opening assembly 217 and the pipetting module 218 are driven by the Z-axis lead screw 216 to achieve independent or coordinated movement in the Z direction.

[0028] It should be noted that the multi-axis motion mechanism includes an X-axis drive assembly, a Y-axis drive assembly, and a Z-axis drive assembly.

[0029] In some embodiments, the X-axis drive assembly includes an X-axis guide rail 21, an X-axis first slider 22, an X-axis motor 23, an X-axis motor mounting plate 24, and an X-axis lead screw 25. Specifically, the X-axis guide rail 21 and the X-axis motor mounting plate 24 are fixed to a base plate. The X-axis motor 23 is fixed to the X-axis motor mounting plate 24. The Y-axis support plate 26 is fixed to the X-axis first slider 22. The X-axis motor 23 drives the Y-axis support plate 26 to move in the X direction via the X-axis lead screw 25. It should be noted that the Y-axis support plate 26 serves as the mounting base for the Y-axis drive assembly, and its displacement along the X-axis enables a wide range of movement of the entire pipetting module 2 in the X direction.

[0030] In some embodiments, the Y-axis drive assembly includes a Y-axis fixing plate 27, a Y-axis guide rail 28, a Y-axis slider 29, a Y-axis motor fixing plate 210, a Y-axis motor 211, and a Y-axis lead screw 212. Specifically, the Y-axis fixing plate 27 and the Y-axis guide rail 28 are fixed to the Y-axis support plate 26. The Y-axis motor 211 is fixed to the Y-axis slider 29. The Y-axis motor 211 drives the Y-axis motor fixing plate 210 to move along the Y-axis guide rail 28 in the Y-axis direction by driving the Y-axis lead screw 212. It should be noted that the displacement of the Y-axis motor fixing plate 210 can simultaneously drive the Z-axis drive assembly, the cap opening assembly 217, and the pipetting module 218 mounted on it to be precisely positioned along the Y-axis, thereby aligning them with the different arrangement positions of the reagent kit.

[0031] In some embodiments, the Z-axis drive assembly includes a Z-axis support plate 213, a Z-axis fixing plate 214, a Z-axis guide rail 215, and a Z-axis lead screw 216. Specifically, the Z-axis support plate 213 is fixed to the Y-axis motor fixing plate 210 and moves synchronously with it in the Y direction. The Z-axis fixing plate 214 and the Z-axis guide rail 215 are fixed to the Z-axis support plate 213. The cap opening assembly 217 and the pipetting module 218 are slidably connected to the Z-axis guide rail 215, and are driven in the Z direction by the Z-axis lead screw 216.

[0032] It should be noted that the vertical movements of the cap opening assembly 217 and the pipetting module 218 can be independent of each other or controlled and coordinated. For example, when performing the cap opening action, the cap opening assembly 217 is driven downward by the Z-axis drive assembly, while the pipetting module 218 can be maintained at a safe height; after the cap is opened, the pipetting module 218 descends to perform liquid retrieval or pipetting operations.

[0033] It should be noted that the specific driving method of the above-mentioned multi-axis motion mechanism includes, but is not limited to, lead screw drive, synchronous belt drive, or linear motor drive, as long as it can achieve the positioning of the pipetting module 218 and the cap opening assembly 217 in three-dimensional space. Specifically, the arrangement order of the sliders, guide rails, and motors of the multi-axis motion mechanism can be flexibly adjusted according to the compactness requirements of the equipment space, and should not be construed as the sole limitation on the scope of protection of this application.

[0034] Optionally, the multi-axis motion mechanism drives the cap-opening assembly 217 to perform a downward cap-opening action. The cap-opening assembly 217 abuts against the flip cap of the reagent kit, causing the flip cap to first flip to a half-open or slightly open state, and then move to a fully open state. Subsequently, the multi-axis motion mechanism drives the pipetting module 218 to enter the opened reagent kit to perform subsequent experimental actions.

[0035] Optionally, the lid opening assembly 217 includes an abutment for performing a press-to-open mechanism. Specifically, in some embodiments, such as Figure 8 and Figure 9 As shown, when the abutment performs the pressing and opening action, the abutment abuts against the flip cover 52 of the reagent kit 5, so that the flip cover 52 first flips to a half-open or slightly open state, and then moves to a fully open state.

[0036] It should be noted that the semi-open or slightly open state refers to the state in which the flip cover 52 is partially opened to release the internal pressure difference of the reagent kit 5. Specifically, the abutment is triggered by pressing the lower part of the flip cover 52 and simultaneously abuts the upper part of the flip cover 52 to limit its opening range, thereby preventing liquid splashing. Optionally, the specific structure of the abutment can be a combination of multiple rods with a fixed length relationship, or it can be an abutment component with an independently adjustable telescopic length.

[0037] In some embodiments, such as Figure 2 , Figure 3 and Figure 4 As shown, the fully automated sequencing library preparation instrument provided in this application includes a frame. Specifically, the frame includes a base plate 4 and a housing 1 fixed on the base plate 4. The housing 1 includes a door 11, a front housing 12, a rear housing 13, a left side housing 14, a right side housing 15, an upper housing 16, a display 17, an emergency stop switch 18, a power filter 19, a filtration system 110, a door magnetic latch 111, a door magnetic plate 112, an indicator light 113, and an ultraviolet disinfection lamp 114.

[0038] Specifically, the front shell 12, rear shell 13, left side shell 14, right side shell 15, and upper shell 16 are interconnected to form the external outline structure of the device body. In some embodiments, this external outline structure is used to improve the aesthetics of the device and to achieve dust and water resistance, thereby providing a controlled working environment for the internal pipetting and temperature transfer modules.

[0039] In some embodiments, the compartment door 11 is disposed on the front housing 12. Specifically, a magnetic plate 112 is fixed to the compartment door 11, and a magnetic latch 111 is fixed to the front housing 12. The magnetic latch 111 and the magnetic plate 112 engage magnetically to open or close the compartment door 11. It should be noted that the specific installation position and magnetic strength of the magnetic adsorption assembly can be adjusted according to the weight of the compartment door 11 to ensure closure stability.

[0040] Specifically, the display 17 is fixed to the front housing 12. In some embodiments, the operator completes the operation settings through the display 17, thereby controlling the device to complete the required experiment. Meanwhile, an indicator light 113 is fixed on the front housing 12 to determine if any abnormalities occur and to provide feedback on the work progress. It should be noted that the display 17 can also be a touchscreen integrated into the device, or it can be a standalone terminal connected to the device via a communication interface.

[0041] Specifically, the ultraviolet disinfection lamps 114 are fixed on the left side shell 14 and the right side shell 15, respectively. Optionally, the ultraviolet disinfection lamps 114 are ozone-free disinfection lamps, used to activate the sterilization mode after the work is completed to prevent cross-contamination of samples. Furthermore, the emergency stop switch 18 is fixed on the right side shell 15. When the equipment experiences alarms or errors, the emergency stop switch 18 is used to immediately cut off the power to the entire machine to avoid equipment damage or personal injury.

[0042] In some embodiments, the filtration system 110 is disposed on the rear housing 13. Specifically, the filtration system 110 includes a cooling fan 1101, a high-efficiency filter 1102, and a filter screen 1103, to prevent gases generated inside the device during ventilation and heat dissipation from being directly discharged into the external environment. In addition, a power filter 19 is fixed on the rear housing 13 to provide power supply and electrical filtering functions for the entire device.

[0043] It should be noted that the connection between the outer casing 1 and the base plate 4 can be achieved by screw fixing, welding, or other structural connections. Specifically, the layout of the various components inside the outer casing 1 (such as the display 17, emergency stop switch 18, etc.) can be optimized according to the overall compactness requirements of the device, and should not be construed as the sole limitation on the scope of protection of this application.

[0044] In some embodiments, such as Figure 7 As shown, the pipetting module 218 includes a pipette 2185. In some embodiments, by controlling the spatial coordinate position and movement of the pipette 2185, the pipetting module 218 can achieve automated liquid collection, pipetting, and mixing. It should be noted that the specific parameters of the target actions can be preset according to the specific process requirements of nucleic acid extraction or library construction.

[0045] Specifically, the pipetting module 218 includes a Z-axis second slider 2181, a Z-axis second motor 2182, a Z-axis second motor fixing plate 2183, a pipette connecting plate 2184, and a pipette 2185.

[0046] Specifically, the Z-axis second motor fixing plate 2183 is fixed to the Z-axis second slider 2181. The Z-axis second motor 2182 and the pipette connecting plate 2184 are fixed to the Z-axis second motor fixing plate 2183. The pipette 2185 is fixed to the pipette connecting plate 2184. It should be noted that by driving the Z-axis second slider 2181 along the Z-axis guide rail 215 through the multi-axis motion mechanism, the entire pipetting module 218 can achieve vertical displacement.

[0047] In some embodiments, by controlling the position and internal mechanism of the pipette 2185, the target actions such as pipetting, liquid collection, and mixing can be completed during the operation. Specifically, liquid collection and pipetting involve the pipetting module 218 being positioned to the designated reagent compartment under the drive of the multi-axis motion mechanism, and the pipette 2185 drawing a preset volume of liquid and transferring it to the target reaction well. Mixing involves the pipette 2185 performing repeated liquid aspiration and dispensing actions within the target reaction well to achieve thorough mixing of different reagent components.

[0048] It should be noted that the spatial coordinates of the pipette 2185 are determined collaboratively by the X, Y, and Z-axis drive components of the multi-axis motion mechanism. Specifically, the Z-axis second motor 2182 can be used to achieve precise control of the piston stroke inside the pipette 2185 to ensure liquid dispensing accuracy. Optionally, the specific parameters of the target action (such as dispensing frequency, dispensing volume, and liquid level detection height) can be preset on the display 17 according to the specific process requirements of nucleic acid extraction or library construction.

[0049] It should be noted that the structural layout and component connection method of the pipetting module 218 can be optimized according to the space compactness requirements of the whole machine. Specifically, the pipette 2185 can be adapted to pipette tips with different volume ranges, depending on the reagent volume range required for the experiment, and should not be construed as the sole limitation on the scope of protection of this application.

[0050] In some embodiments, such as Figure 10 , Figure 11 and Figure 12 As shown, the fully automated sequencing library preparation instrument includes an incubation transport module 3. Specifically, the incubation transport module 3 includes an X-axis second slider 31, an incubation module base plate 32, a front support plate 33, a left support plate 34, a right support plate 35, a reagent kit fixing plate 36, an incubation module 37, and a magnetic suction module 38.

[0051] Specifically, the incubation module base plate 32 is fixed to the X-axis second slider 31. In some embodiments, the incubation transfer module 3 can move along the X-direction via the X-axis second slider 31, thereby automating the placement and retrieval of the reagent kit 5. It should be noted that this X-direction movement displacement is controlled collaboratively by a multi-axis motion mechanism to ensure that the reagent kit 5 is accurately moved to the pipetting or capping station. Further, the front support plate 33, the left support plate 34, and the right support plate 35 are fixed to the incubation module base plate 32. The reagent kit fixing plate 36 is fixed to the left support plate 34 and the right support plate 35, together forming the overall frame structure of the incubation transfer module 3.

[0052] In some embodiments, the incubation transfer module 3 integrates an incubation module 37 for controlling the reaction temperature. Specifically, the incubation module 37 includes an incubation tank 371, a heat sink 372, a heat dissipation support frame 373, a cooling fan 374, and a Peltier 375. The heat dissipation support frame 373 is fixed to the base plate 32 of the incubation module, and the heat sink 372 is fixed to the heat dissipation support frame 373. The incubation tank 371 is fixed to the heat sink 372 by the Peltier 375 and thermal grease. It should be noted that when the Peltier 375 is heated in a controlled manner, the heat is conducted to the incubation tank 371 through the thermal grease, thereby performing an incubation operation on the reagent kit 5 placed in the incubation tank 371. The cooling fan 374 is fixed to the heat sink 372 to accelerate the cooling process after heating, thereby completing the heating and cooling cycle control of the incubation module 37.

[0053] In some embodiments, the incubation transfer module 3 also integrates a magnetic adsorption module 38 for magnetic bead separation. Specifically, the magnetic adsorption module 38 includes a magnet 381, a magnet connector 382, ​​a push rod 383, and a push rod fixing seat 384. The magnet 381 is fixed to the push rod 383 via the magnet connector 382, ​​and the push rod 383 is fixed to the incubation module base plate 32 via the push rod fixing seat 384. By controlling the extension and retraction of the push rod 383, the magnet 381 is driven to move up and down in the Z direction, thereby realizing the adsorption or release of magnetic beads in the reaction wells of the reagent kit 5 by the magnet 381. It should be noted that through the adsorption and separation of magnetic beads, the transfer and purification of liquid reagents can be completed in conjunction with the pipetting module.

[0054] In addition, such as Figure 13 As shown, in one optional embodiment, the fully automated sequencing library preparation instrument supports a multi-machine parallel operation mode. It should be noted that parallel operation of multiple devices enables the parallel processing of multiple batches of samples, thereby significantly improving sample processing throughput and meeting the needs of large-scale library construction.

[0055] It should be noted that the connection method of each support plate in the incubation and transmission module 3 and the driving stroke of the magnetic suction module 38 can be adapted to the specifications of the reagent kit 5. Specifically, the temperature control accuracy of the Peltier 375 and the magnetic field strength of the magnet 381 can be optimized according to the specific biochemical experimental requirements, and should not be construed as the sole limitation on the scope of protection of this application.

[0056] The fully automated sequencing library preparation instrument provided in this application, through the step-by-step stroke control logic of the capping component 217, in conjunction with the cooperation of the multi-axis motion mechanism and the pipetting module 218, can significantly reduce the risk of contamination during library preparation and improve experimental efficiency. It should be noted that the above general description aims to illustrate the basic architecture of this application; the specific structure and control flow of each functional component will be further elaborated in subsequent embodiments.

[0057] In some embodiments, the fully automated sequencing library preparation instrument includes a cap-opening assembly 217 disposed on a base plate 4. Specifically, as Figure 6 As shown, the cover opening assembly 217 includes a Z-axis first slider 2171, a Z-axis first motor 2172, a Z-axis first motor fixing plate 2173, a fixing base 2174, and an abutment member.

[0058] Specifically, the Z-axis first motor fixing plate 2173 is fixed to the Z-axis first slider 2171. The Z-axis first motor 2172 and the fixing seat 2174 are fixed to the Z-axis first motor fixing plate 2173. The abutment extends along the downward pressing opening direction of the fixing seat 2174. It should be noted that by driving the Z-axis first slider 2171 to move along the Z-axis guide rail 215 through the multi-axis motion mechanism, the entire opening assembly 217 can perform a vertical lifting action relative to the base plate 4.

[0059] In some embodiments, the abutment includes a first abutment portion and a second abutment portion. Specifically, refer to... Figure 14 The first abutment part is used to press down the upper flip part 522 and the lower flip part 521 of the reagent kit cover when it is in the closed state, so that the cover is opened to a half-open or slightly open state; the second abutment part is used to press down the lower flip part 521 of the reagent kit cover when it is in the half-open or slightly open state, so that the cover is opened to a fully open state.

[0060] In some embodiments, such as Figure 6 As shown, the abutment component is implemented using a multi-bar combination structure. Specifically, the first abutment part includes a first bar 2176 and a second bar 2175; the second abutment part includes a third bar 2177.

[0061] It should be noted that, in order to meet the logical requirement of step-by-step opening, the length of the third rod 2177 is greater than the length of the first rod 2176, and the length of the first rod 2176 is greater than the length of the second rod 2175.

[0062] Specifically, refer to Figure 8 , Figure 14During the semi-opening or slightly-opening phase, the multi-axis motion mechanism drives the lid opening assembly 217 to perform the first downward stroke. At this time, the first rod 2176, which is of medium length, first contacts and presses down on the lower flip portion 521 of the reagent kit lid, inducing the lid to open upward. Subsequently, the second rod 2175, which is the shortest, presses down and abuts against the upper flip portion 522, which pops up synchronously with the lid. In this state, the first rod 2176 and the second rod 2175 work together to flip the lid to a semi-open or slightly-open state, thereby slowly releasing the internal pressure of the reagent kit. During this stroke, the third rod 2177 no longer mechanically interferes with the reagent kit 5. It should be noted that the absence of mechanical interference here means that the third rod 2177 is in free space during the pressing process and will not contact any part of the reagent kit 5, thereby avoiding prematurely pressing the lid to the fully open state due to the excessive length of the third rod 2177.

[0063] Specifically, such as Figure 9 As shown, during the fully open phase, the multi-axis motion mechanism drives the opening assembly 217 to switch its horizontal coordinates, aligning the longest third rod 2177 with the lower flip portion 521 and executing the second downward stroke. Due to the length advantage of the third rod 2177, it presses the lower flip portion 521 to its limit position, causing the flip to move to the fully open state. During this stroke, the first rod 2176 and the second rod 2175 no longer mechanically interfere with the reagent kit 5. It should be noted that the absence of mechanical interference here means that during the downward pressing of the third rod 2177, the first rod 2176 and the second rod 2175 do not obstruct the upward flipping trajectory of the upper flip portion 522, thereby ensuring that the flip can move smoothly to the fully open state and avoiding collision damage between the rods and the flip.

[0064] In some alternative embodiments, the abutment of the opening assembly 217 implements a step-by-step opening logic through a specific geometric surface profile. Specifically, the first abutment is a first contact surface with a first geometric configuration disposed at the bottom end of the abutment; the second abutment is a second contact surface with a second geometric configuration disposed at the bottom end of the abutment.

[0065] Specifically, the orthographic projection area of ​​the first contact surface on the horizontal plane of the base plate 4 at least partially covers the orthographic projection area of ​​the lower flip portion 521 of the reagent kit cover on the horizontal plane of the base plate 4, and at the same time at least partially covers the orthographic projection area of ​​the upper flip portion 522 of the cover on the horizontal plane of the base plate 4.

[0066] During the semi-opening or slightly-opening operation, the multi-axis motion mechanism drives the opening assembly 217 downward along the Z-axis. Specifically, the portion of the first contact surface corresponding to the downward-flipping part 521 first contacts and presses downward against the downward-flipping part 521, inducing the flip cover to rotate upward around its axis of rotation and open. At the same time, since the projection area of ​​the first contact surface also covers the movement path of the upward-flipping part 522, the portion of the first contact surface corresponding to the upward-flipping part 522 acts as a mechanical limiting surface, abutting against the upward-flipping part 522, which rotates synchronously with the flip cover and springs upward. It should be noted that through the power output to the downward-flipping part 521 and the resistance limiting to the upward-flipping part 522 by the first contact surface in the same displacement stroke, the flip cover flips to the semi-opening or slightly-opening state, thereby achieving the balance of air pressure inside and outside the reagent kit and preventing liquid splashing.

[0067] Specifically, the orthographic projection area of ​​the second contact surface on the horizontal plane of the base plate 4 only covers the orthographic projection area of ​​the lowered portion 521 on the horizontal plane of the base plate 4, and avoids the rotation path of the uppered portion 522 in the vertical direction. It should be noted that the first contact surface and the second contact surface at the bottom end of the abutment can be a stepped structure, a sloped structure, or a discontinuous protrusion structure.

[0068] During the fully opening operation, the multi-axis motion mechanism drives the opening assembly 217 to switch coordinates in space, aligning the second contact surface with the lowering portion 521, while moving the first contact surface above the uppering portion 522. Subsequently, the opening assembly 217 performs a second pressing stroke, pressing the lowering portion 521 to a preset limit position by the second contact surface. It should be noted that during the pressing process of the second contact surface, since the remaining parts of the abutment no longer have physical contact or interference with the uppering portion 522, the flip cover 52 can smoothly move to the fully open state.

[0069] It should be noted that this design based on the contact surface configuration allows the first and second contact portions to be integrated into the same integrally molded part. Specifically, the surface of the contact surface can be processed with anti-slip texture or covered with soft material to increase the gripping force when in contact with the flip-down portion 521. In addition, the height difference or horizontal spacing between the first and second contact surfaces can be customized according to the rotation radius of the reagent kit flip-top 52 produced by different manufacturers.

[0070] It should be noted that the components of the aforementioned abutment (first rod 2176, second rod 2175, and third rod 2177) can be detachably fixed to the mounting base 2174. Specifically, the Z-axis first motor 2172 can be used to finely adjust the height or downward pressure of the mounting base 2174 to adapt to reagent kit caps 52 made of different materials and with different opening force requirements.

[0071] In some embodiments, the fully automated sequencing library preparation instrument includes a capping assembly 217 disposed on a base plate 4. Specifically, the capping assembly 217 includes a mounting base and an abutment mounted on the mounting base. In this embodiment, the abutment includes a third abutment portion and a fourth abutment portion with independently adjustable telescopic lengths.

[0072] Specifically, both the third and fourth abutment portions are connected to the drive mechanism. It should be noted that the drive mechanism may include a micro servo motor, an electromagnetic actuator, or a pneumatic telescopic element to drive the third and fourth abutment portions to extend different lengths relative to the fixed base. Through the coordination of the multi-axis motion mechanism and the drive mechanism, the opening assembly 217 can change the opening state of the reagent kit 5's flip-top by switching the extension and retraction states of the abutment portions.

[0073] Specifically, during the semi-opening or slightly-opening stage, the multi-axis motion mechanism drives the cap-opening assembly 217 to position above the reagent kit 5. The third abutment extends by a first extension to abut the lower flap 521 of the cap, while the fourth abutment extends by a second extension to abut the upper flap 522. It should be noted that the first extension is greater than the second extension at this time. In this state, the third abutment presses downwards on the lower flap 521 to induce opening, while the fourth abutment limits the upward flap 522 as it springs up. Through the cooperation of both, the cap flips to a semi-open or slightly-open state to release internal pressure in the reagent kit 5 and prevent liquid splashing.

[0074] Specifically, during the fully opening phase, the multi-axis motion mechanism maintains the position of the opening assembly 217 or performs fine adjustments. The third abutment extends by a third elongation to continue abutting the lower flip-up portion 521, while simultaneously driving the fourth abutment to retract upward to disengage from the upper flip-up portion 522.

[0075] It should be noted that since the fourth abutment no longer interferes with the movement path of the flip cover, the third abutment presses the lower flip part 521 to the limit position by a greater extension, thereby performing the operation of opening the flip cover to the fully open state.

[0076] It should be noted that in the operation logic of this embodiment, the third elongation, the first elongation, and the second elongation decrease sequentially. Specifically, this dynamic switching of the length gradient allows the lid opening component 217 to complete the entire lid opening process without performing complex avoidance actions in the horizontal direction, simply by adjusting the length in the vertical direction.

[0077] It should be noted that the spacing between the third and fourth abutting parts on the fixing base can be preset according to the structural dimensions of the flip-top of the reagent kit 5. In some optional embodiments, pressure sensors can be provided at the ends of the third and fourth abutting parts to provide real-time feedback on the abutting force, ensuring that the reagent kit 5 is not damaged during half-opening, slightly opening, or fully opening operations.

[0078] Example 2 According to one aspect of this application, a control method for a fully automated sequencing library preparation instrument is provided, applied to the fully automated sequencing library preparation instrument of the foregoing embodiments. This fully automated sequencing library preparation instrument, during the process of nucleic acid extraction or library construction, such as… Figure 15 As shown, the control method includes the following steps: S1. Perform positioning preparation actions before opening the lid.

[0079] In some embodiments, the multi-axis motion mechanism is controlled to drive the opening assembly to move directly above the reagent kit, such that the first abutment portion of the abutment is aligned with the lower flip portion 521 and the upper flip portion 522 of the flip cover. It should be noted that the accuracy of this positioning action is ensured by the X-axis and Y-axis pulse control of the multi-axis motion mechanism.

[0080] S2. Control the opening component to perform the first pressing action, so that the opening component simultaneously abuts against the lower and upper flip parts of the reagent kit cover, so that the cover is pressed down to open to a half-open or slightly open state.

[0081] Specifically, the Z-axis drive mechanism of the multi-axis motion mechanism or the cover opening assembly drives the abutment to move downward along the Z-axis. During this process, the first abutment triggers the cover to rotate by pressing down the flip-up part 521, and simultaneously abuts against the flip-up part 522 to limit the opening stroke of the cover.

[0082] It should be noted that the technical effect of this step is to physically limit the flip-top to a semi-open or slightly open state, thereby allowing the pressure difference inside the reagent kit caused by biochemical reactions or incubation to be slowly released. Specifically, this step-by-step action can effectively balance the internal and external pressures and prevent the risk of liquid splashing caused by the flip-top popping up instantly.

[0083] S3. Control the opening assembly to perform a second pressing action, so that the flip cover moves from a half-open or slightly open state to a fully open state.

[0084] Specifically, after a preset time (e.g., 0.5 to 2 seconds) when the flip cover is flipped to a half-open or slightly open state, the control of the opening component switches the pressing logic.

[0085] S4. Perform the target action for the current nucleic acid extraction or library construction.

[0086] Specifically, after the flip-top is fully open, the multi-axis motion mechanism drives the pipetting module 218 into the reaction well of the reagent kit. Target actions include, but are not limited to, liquid collection, pipetting, and mixing. It should be noted that the specific parameters of the target actions (such as liquid level detection depth, aspiration rate, and mixing frequency) can be preset in the system according to experimental requirements.

[0087] S5. Control the lid opening component to close the flip cover.

[0088] After the target action is completed, the control pipetting module 218 is ejected from the reagent kit. Specifically, the control cap opening assembly or multi-axis motion mechanism performs the cap closing action, resetting the cap to the closed state to prevent external impurities from contaminating the sample or reagents from evaporating.

[0089] It should be noted that each step in the above control method can be executed cyclically according to experimental requirements. For example, when processing multiple rows of reagent wells, the system can sequentially execute the above steps S2 to S5 for each well. Specifically, the control method provided in this application achieves mechanical graded control of the opening amplitude through the timing coordination of S2 and S3.

[0090] It should be noted that this control method is not only applicable to the aforementioned multi-bar opening assembly, but also to opening assemblies with telescopic contact parts or specific contact surface configurations. This application, through this control method, ensures the safety and standardization of the automated library construction process at the underlying algorithmic logic level, effectively avoiding the risks of cross-contamination and splashing caused by manual operation and traditional one-stop opening solutions.

[0091] In a specific embodiment, such as Figure 6 , Figure 8 and Figure 9 As shown, the cover opening assembly 217 includes a first rod 2176, a second rod 2175, and a third rod 2177. The length of the third rod 2177 is greater than the length of the first rod 2176, and the length of the first rod 2176 is greater than the length of the second rod 2175.

[0092] Based on this hardware, the control method controls the opening component to perform the first pressing action. The opening component simultaneously abuts against the lower flip part 521 and the upper flip part 522 of the reagent kit cap, so that the cap is pressed open to a half-open or slightly open state. Step S2 specifically includes the following steps: S211, Control the multi-axis motion mechanism to drive the cover opening assembly 217 to move to the first cover opening position.

[0093] Specifically, this step aims to align the first lever 2176 with the lower part 521 of the reagent kit cap, while aligning the second lever 2175 with the upper part 522 of the reagent kit cap. It should be noted that the determination of the first opening position relies on preset reagent kit coordinates to ensure that the two levers accurately act on the target area during the subsequent downward pressure stroke.

[0094] S212, Control the opening assembly 217 to perform the first pressing action.

[0095] Specifically, the multi-axis motion mechanism or Z-axis drive assembly drives the fixed base downward, so that the first rod 2176 abuts against the lower flip part 521 to provide opening force, while the second rod 2175 abuts against the upper flip part 522 to limit the rotation amplitude, thereby pressing the flip cover down to open it to a half-open or slightly open state.

[0096] Furthermore, step S3, which controls the opening assembly to perform a second pressing action so that the flip cover moves from a half-open or slightly open state to a fully open state, specifically includes the following steps: S311, Control the multi-axis motion mechanism to drive the cover opening assembly 217 to move to the second cover opening position.

[0097] Specifically, this step involves aligning the third rod 2177 with the lowered section 521 via horizontal displacement. It should be noted that during the movement to the second open position, the first rod 2176 and the second rod 2175 simultaneously move out of the rotation path of the flip cover to avoid mechanical interference in subsequent movements.

[0098] S312, Control the opening assembly 217 to perform the second pressing action.

[0099] Specifically, the third lever 2177 abuts against the lowered section 521 and presses it to its limit position, causing the flap to open fully. Because the third lever 2177 has the maximum length, this stroke ensures that the flap opens completely without being obstructed by other levers.

[0100] In a specific embodiment, such as Figure 14 As shown, the reagent kit flap 52 includes a lower flap 521, an upper flap 522, and a groove structure 523. The groove structure 523 is disposed between the lower flap 521 and the upper flap 522 and extends along the length of the flap. Specifically, the groove structure 523 is used to provide guidance and force support for the abutment during the closing stroke.

[0101] Based on this structure, the aforementioned Figure 15 In the process, step S5, which controls the opening assembly to close the flip cover, specifically includes the following sub-steps: S511, Control the lid opening assembly to perform an upward movement at the second lid opening position.

[0102] Specifically, refer to Figure 14 The Z-axis mechanism drives the third rod 2177 upward, raising its bottom to a position aligned with or slightly above the top surface of the flip cover 52 when it is fully closed. This height positioning is to ensure that the third rod 2177 can accurately engage the slotted structure 523 during subsequent horizontal movement without causing a hard vertical collision.

[0103] S512, Control the multi-axis motion mechanism to drive the third rod to move to the closed position located on the surface of the upper flip part, so as to close the flip cover.

[0104] Specifically, refer to Figure 14 The multi-axis motion mechanism drives the third rod 2177 to move horizontally in the direction pointing towards the rotation axis. During this stroke, the bottom end of the third rod 2177 enters and slides along the groove structure 523, thereby using the side wall of the groove structure 523 to horizontally limit the third rod 2177. It should be noted that through the cooperation of the third rod 2177 and the groove structure 523, it can be ensured that the thrust always acts on the central axis of the flip cover 52, preventing the flip cover 52 from lateral displacement or jamming during the closing process, and finally pushing the flip cover 52 from the fully open state back to the closed state.

[0105] It should be noted that the depth and width of the groove structure 523 are adapted to the diameter of the third rod 2177. Specifically, by providing the groove structure 523 on the surface of the flip cover 52, this application not only automates the opening action, but also significantly reduces the extreme dependence on the motion accuracy of the multi-axis motion mechanism by utilizing the physical guidance of the mechanical structure, thereby improving the closing stability of the fully automated sequencing library preparation instrument during long-term operation.

[0106] It should be noted that the specific stroke parameters (such as pushing speed and horizontal displacement) of the above-mentioned closing action can be preset according to the material resistance of the flip cover 52. In some optional embodiments, after the third rod 2177 enters the groove structure 523, it can cooperate with a small Z-axis downward pressure component to ensure that the flip cover 52 can be reliably fastened to the reagent kit body.

[0107] In one specific embodiment, the abutment of the opening assembly 217 includes a third abutment portion and a fourth abutment portion with independently adjustable extension lengths. This fully automated sequencing library preparation instrument control method switches the opening state of the flip cover 52 by adjusting the extension lengths of the third and fourth abutment portions.

[0108] Before performing the opening action, the control method first includes the following alignment steps: S120, Control the opening assembly to move so as to align with the flip cover.

[0109] Specifically, by controlling the multi-axis motion mechanism, the third abutment portion is aligned with the lower flip portion 521 of the reagent kit cover, and the fourth abutment portion is aligned with the upper flip portion 522 of the reagent kit cover. It should be noted that this alignment step ensures that the subsequent telescopic action can be accurately applied to the designated force point of the cover 52.

[0110] Step S2, which controls the opening assembly 217 to perform the first pressing action and opens the cover 52 to a half-open or slightly open state by simultaneously abutting the lower flip part 521 and the upper flip part 522, specifically includes: S220, control the third abutting part to extend by a first extension to abut the lower flipping part, and simultaneously control the fourth abutting part to extend by a second extension to abut the upper flipping part.

[0111] During this process, the first extension is greater than the second extension, thereby driving the flip cover 52 to press down and open to a half-open or slightly open state. It should be noted that, through the height difference between the third and fourth abutment parts, the flip cover 52 is restricted within a predetermined range at the moment of opening, thereby achieving a slow release of the internal air pressure of the reagent kit.

[0112] Step S3, which controls the opening assembly 217 to perform the second pressing action, causing the flip cover 52 to move from a half-open or slightly open state to a fully open state, specifically includes: S320, Control the third abutment portion to extend from the first elongation to the third elongation, and control the fourth abutment portion to disengage from the upturned portion.

[0113] Specifically, the third abutment portion is driven to extend downwards to the third elongation, while the fourth abutment portion is controlled to retract upwards, so that the flip cover 52 is pressed down and opened to the fully open state. It should be noted that in this step, the third elongation, the first elongation, and the second elongation decrease sequentially. Because the fourth abutment portion disengages in time, the flip cover 52 is able to complete the maximum angle of rotation under the drive of the third abutment portion.

[0114] After completing the target action step S4, step S5, which controls the opening assembly 217 to close the flip cover 52, specifically includes the following sub-steps: S521, Control the opening assembly to perform an upward action and switch the third abutment part to the third extension amount.

[0115] Specifically, the Z-axis mechanism is controlled to retract upwards, aligning the bottom height of the third abutment with the top surface height of the flip cover 52 in the closed state. It should be noted that maintaining the third abutment at a relatively long third extension helps provide better lever arm support during the closing process.

[0116] S522, Control the multi-axis motion mechanism to drive the third abutment part to the closed position located on the surface of the upper flip part, so as to close the flip cover.

[0117] Specifically, the horizontal thrust of the multi-axis motion mechanism causes the third abutment part to slide along the surface of the flip cover 52 and push it back to the closed state. Combined with... Figure 15 As shown, during the sliding process, the bottom end of the third abutment part can enter the groove structure 523. Through the physical limiting and guiding effect of the groove structure 523, the thrust is always applied to the effective area of ​​the flip cover 52, thereby achieving precise closing of the cover.

[0118] It should be noted that the extension and retraction control of the third and fourth contact parts can be achieved using a servo motor or a precision push rod. Specifically, by using the system's preset elongation parameters for each stage, this method can adapt to experimental requirements with different opening amplitudes and can achieve a dynamic transition from half-open or slightly open to fully open without frequently switching the horizontal position.

[0119] The control method provided in this embodiment utilizes the independent telescopic characteristics of the contact part to simplify the automated operation path. While improving the safety of opening the lid, it further enhances the fully automated sequencing library preparation instrument's ability to operate complex consumables.

[0120] In a specific embodiment, during nucleic acid extraction or library construction, step S4 is executed collaboratively by the pipetting module 218, the incubation and transfer module 3, and the capping component 217. Specifically, step S4 comprises a sequence of multiple biochemical process sub-steps, and during each sub-process, the system cyclically calls steps S2, S3, and S5 to open the cap 52 before liquid addition or transfer operations and to promptly close the cap 52 during incubation, magnetic suction, or standby states, thereby preventing reagent evaporation and cross-contamination between samples.

[0121] When the fully automated sequencing library preparation instrument performs nucleic acid extraction, step S4 specifically includes the following sub-step sequence: In step one, the system controls the multi-axis motion mechanism to drive the capping component 217 to execute steps S2 and S3 to open the cap 52 to the fully open state. Then, the system controls the pipetting module 218 to add 200 μL of sample, 300 μL of lysis buffer, 20 μL of PK buffer, and 15 μL of magnetic bead solution to reaction position 1 in sequence and mix them. After the action is completed, the system controls the capping component 217 to execute step S5 to close the cap 52, and the incubation module 37 performs heating incubation. The supernatant is discarded with the assistance of the magnetic suction module 38. In step two, steps S2 and S3 are repeated to open the cap 52, and the pipetting module 218 adds 200 μL of sample, 300 μL of lysis buffer, 20 μL of PK buffer, and 15 μL of magnetic bead solution to reaction position 1 and mixes them. In step 1, add 480 μL of protein removal solution and mix well. After the operation is completed, proceed to step S5, close the flap 52, and discard the supernatant. In step 3, open the flap 52 and add 750 μL of washing solution. After the operation is completed, close the flap 52 and discard the supernatant. In step 4, open the flap 52 and add 33 μL of elution solution and mix well. After closing the flap 52, heat to 56°C using the incubation module 37 for incubation and elution. Finally, discard the magnetic beads using the magnetic adsorption module 38 and retain the elution solution. In step 5, open the flap 52 and add 50 μL of fragmentation reagent and mix well. After closing the flap 52, heat to 32°C using the incubation module 37 for incubation. In step 6, perform final quality control analysis on the extracted product.

[0122] When the fully automated sequencing library construction instrument performs library construction, step S4 specifically includes the following sub-step sequences: In step one, the system opens the flap 52 of reaction position 2, and the pipetting module 218 adds 50 μL of fragmented DNA, 10 μL of reagent 1, and 5 μL of enzyme 1 and mixes them. After closing the flap 52 in step S5, the incubation module 37 performs end-repair incubation at 20°C and 65°C sequentially. In step two, the flap 52 is opened and 5 μL of reagent 2, 26 μL of reagent 3, and 4 μL of enzyme 2 are added and mixed. After closing the flap 52, the mixture is incubated at 20°C to complete the adapter ligation operation. In step three, the liquid is transferred to reaction position 3, and the flap 52 is opened. 2. Add 80 μL and 50 μL of magnetic bead solution and mix well. After closing the flip-top 52, the supernatant is discarded and ethanol rinsing is performed with the assistance of the magnetic suction module 38. In step four, open the flip-top 52 and add 20 μL of reagent 4 for elution. After the elution is completed, close the flip-top 52. In step five, transfer the supernatant to reaction site 4, open the flip-top 52 and add 5 μL of reagent 5 and 25 μL of enzyme 3. After closing the flip-top 52, the incubation module 37 performs PCR amplification cycles including temperature variation nodes of 60°C, 72°C and 98°C. In subsequent steps six to eight, repeat the logic of opening and closing the flip-top 52 to complete the secondary magnetic bead purification, reagent 4 elution and final library quality control test.

[0123] To ensure the mechanical reliability of the aforementioned high-frequency opening and closing cycle, in each step S5 of closing the flip cover 52, the system drives the third rod 2177 to move to the closed position located on the surface of the upper flip portion 522. Specifically, in conjunction with Figure 14 The bottom end of the third rod 2177 enters and slides along the groove structure 523 provided on the surface of the flip cover 52. The physical limiting and guiding effect of the groove structure 523 in the horizontal direction is used to offset the slight displacement deviation that may be generated by the multi-axis motion mechanism during high-speed movement, ensuring that the thrust always acts on the central force area of ​​the flip cover 52, thereby achieving precise fastening of the flip cover 52.

[0124] It should be noted that step S4, by embedding step-by-step opening logic between each two adjacent liquid addition or incubation steps, addresses the risk of liquid splashing caused by sudden changes in the pressure difference between the internal air pressure of the kit 5 and the external environment when the lid 52 is opened at 56°C or after PCR high-temperature cycling. This control method clarifies the temporal correlation and logical feedback between the hardware modules (pipette module 218, incubation transfer module 3, lid opening component 217) and biochemical process parameters (reagent volume, temperature control node, magnetic suction timing), ensuring the reproducibility and stability of the entire fully automated sequencing library construction process. It should be noted that the specific reagent volume, incubation temperature, and time parameters in step S4 can be preset according to different nucleic acid extraction kits or library construction strategies, and should not be construed as the sole limitation on the scope of protection of this application.

[0125] Although embodiments of the present invention have been disclosed above, they are not limited to the applications listed in the specification and embodiments. They can be applied to various fields suitable for the present invention. For those skilled in the art, other modifications can be easily made. Therefore, without departing from the general concept defined by the claims and their equivalents, the present invention is not limited to the specific details and illustrations shown and described herein.

Claims

1. A fully automated sequencing library preparation instrument, characterized in that, include: A frame, a pipetting module and an incubation and transfer module disposed within the frame; The incubation transport module is used to carry the reagent kit; The pipetting module includes a multi-axis motion mechanism, a pipetting module, and a cap opening assembly; The multi-axis motion mechanism is connected to the pipetting module and the cap opening assembly, and is used to drive the pipetting module and the cap opening assembly to move relative to the incubation and transfer module; wherein... The lid opening assembly includes an abutment for performing a downward pressing action to open the lid; When the abutment performs the pressing action to open the lid, the abutment abuts against the flip lid of the reagent kit, causing the flip lid to first flip to a half-open or slightly open state, and then move to a fully open state.

2. The fully automated sequencing library preparation instrument according to claim 1, characterized in that, The opening assembly further includes a fixing base; the abutting member extends along the downward opening direction of the fixing base; the abutting member includes a first abutting portion and a second abutting portion; The first abutting part is used to press down the upper and lower flip parts of the flip cover when it is in the closed state, so that the flip cover is opened to a half-open or slightly open state; The second abutting part is used to press down the lower flip part of the cover when it is in a half-open or slightly open state, so that the cover is opened to a fully open state.

3. The fully automated sequencing library preparation instrument according to claim 2, characterized in that, The first abutting part includes a first rod and a second rod, and the second abutting part includes a third rod; The first rod is used to press down the downward flip-up part to trigger opening, and cooperates with the second rod to abut the upward flip-up part to maintain the half-open or slightly open state. The third rod is used to continue pressing down the flip-down part after the half-open or slightly open state, so as to drive the flip cover to the fully open state.

4. The fully automated sequencing library preparation instrument according to claim 2, characterized in that, The first abutting portion and the second abutting portion are different contact surfaces at the bottom end of the abutting member; When the first abutting part performs a downward pressing stroke on the flip cover, it can contact the lower flip part and the upper flip part, so that the flip cover flips to a half-open or slightly open state; When the second abutment part performs a downward pressing stroke on the flip cover, it can contact the flip-up part so that the flip cover is opened to the fully open state.

5. The fully automated sequencing library preparation instrument according to claim 1, characterized in that, The abutting component includes a third abutting part and a fourth abutting part whose extension and retraction lengths can be independently adjusted; the multi-axis motion mechanism drives the third abutting part and the fourth abutting part to extend to different lengths to switch the open state of the flip cover; The third abutment extends by a first elongation to abut the lower flip portion of the cover, while the fourth abutment extends by a second elongation to abut the upper flip portion, thereby causing the cover to first flip to a half-open or slightly open state. The third abutment extends by a third elongation to abut the lower flip portion of the flap, while the fourth abutment disengages from the upper flip portion to open the flap to the fully open state; wherein the third elongation, the first elongation, and the second elongation decrease sequentially.

6. A control method for a fully automated sequencing library preparation instrument, characterized in that, Applied to the fully automated sequencing library preparation instrument as described in claim 1, the fully automated sequencing library preparation instrument includes the following steps during the process of nucleic acid extraction or library construction: The opening assembly is controlled to perform a first pressing action, which simultaneously abuts against the lower and upper flip parts of the reagent kit cap, so that the cap is pressed down to the half-open or slightly open state; the opening assembly is controlled to perform a second pressing action, so that the cap moves from the half-open or slightly open state to the fully open state. Perform the current target action of nucleic acid extraction or library construction; Control the opening component to close the flip cover.

7. The control method according to claim 6, characterized in that, The cover opening assembly includes a first rod, a second rod, and a third rod, wherein the length of the third rod is greater than the length of the first rod, and the length of the first rod is greater than the length of the second rod. The control of the opening component to perform a first pressing action, wherein the opening component simultaneously abuts against the lower and upper flip parts of the reagent kit cap, so that the cap is pressed open to the half-open or slightly open state, specifically includes the following steps: The multi-axis motion mechanism is controlled to drive the opening assembly to move to the first opening position, so that the first rod is aligned with the lower part of the reagent kit cover and the second rod is aligned with the upper part of the reagent kit cover. The opening assembly is controlled to perform a first pressing action, so that the first rod abuts against the lower part of the reagent kit cover and the second rod abuts against the upper part of the reagent kit cover, so that the cover is pressed down to open to the half-open or slightly open state. The step of "controlling the opening assembly to perform a second pressing action so that the flip cover moves from the half-open or slightly open state to the fully open state" specifically includes the following steps: The multi-axis motion mechanism is controlled to drive the opening assembly to move to the second opening position so that the third rod is aligned with the lower part of the reagent kit lid; The opening assembly is controlled to perform a second pressing action, so that the third rod abuts against the lower part of the reagent kit cap, thereby pressing the cap down to the fully open state.

8. The control method according to claim 7, characterized in that, The steps for controlling the opening component to close the flip cover specifically include: The opening assembly is controlled to lift at the second opening position so that the bottom of the third rod is aligned with the top surface of the flip cover when it is closed. The multi-axis motion mechanism is controlled to drive the third rod to move to the closed position located on the surface of the upper flip part, so as to close the flip cover.

9. The control method according to claim 6, characterized in that, The abutment includes a third abutment portion and a fourth abutment portion whose extension and retraction lengths can be independently adjusted; Before the step of controlling the opening component to perform the first pressing action, by having the opening component simultaneously abut against the lower and upper flip parts of the reagent kit cap, so that the cap is pressed open to the half-open or slightly open state, the method further includes the following step: The opening assembly is controlled to move to align with the flip-top, such that the third abutment is aligned with the lower flip-top of the reagent kit flip-top, and the fourth abutment is aligned with the upper flip-top of the reagent kit flip-top. The control of the opening component to perform a first pressing action, wherein the opening component simultaneously abuts against the lower and upper flip parts of the reagent kit cap, so that the cap is pressed open to the half-open or slightly open state, specifically includes the following steps: The third abutting part is controlled to extend by a first extension to abut the lower flip-up part, and the fourth abutting part is simultaneously controlled to extend by a second extension to abut the upper flip-up part, so that the flip cover is pressed down and opened to the half-open or slightly open state; wherein, the first extension is greater than the second extension. The control of the opening assembly to perform a second pressing action, so that the flip cover moves from the half-open or slightly open state to the fully open state, specifically includes the following steps: The third abutment portion is controlled to extend from the first extension amount to the third extension amount, and the fourth abutment portion is controlled to disengage from the flip-up portion, so that the flip cover is pressed down and opened to the fully open state.

10. The control method according to claim 9, characterized in that, The steps for controlling the opening component to close the flip cover specifically include: The opening assembly is controlled to perform an upward movement, and the third abutment is switched to the third extension, so that the bottom height of the third abutment is aligned with the top surface height of the flip cover in the closed state; The multi-axis motion mechanism is controlled to drive the third abutment to move to the closed position located on the surface of the upper flip-up part, so as to close the flip-up cover.

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