Nucleic acid sequencing pretreatment workstation

Abstract

Provided in the present application is a nucleic acid sequencing pretreatment workstation. In the nucleic acid sequencing pretreatment workstation, a nucleic acid extraction system is configured to controllably extract nucleic acid molecules from a biological sample; a library construction system is configured to controllably prepare the nucleic acid molecules into a library which can be used for sequencing on a machine; and a quality control system is configured to controllably measure the concentration of a nucleic acid extraction product and add a normalization reagent to the nucleic acid extraction product on the basis of the measurement result and library construction sample loading parameters, and / or controllably measure the concentration of a library construction product, add a normalization reagent to the library construction product on the basis of the measurement result, and dilute the library construction product to a concentration required for nucleic acid sequencing. By means of the cooperation of several systems, operations such as nucleic acid extraction, nucleic acid extraction product quality control, library construction, and library construction product quality control are implemented, the degree of automation of nucleic acid sequencing pretreatment is improved, and the consistency of nucleic acid sequencing is improved.

Description

Nucleic acid sequencing pre-treatment workstation

[0001] This application claims priority to Chinese Patent Application No. 202411299715.1, filed on September 14, 2024, entitled "Library Construction Workstation and Nucleic Acid Library Construction Method", Chinese Patent Application No. 202422273091.8, filed on September 14, 2024, entitled "Library Construction Workstation", Chinese Patent Application No. 202411719919.6, filed on November 27, 2024, entitled "Nucleic Acid Extraction Instrument and Library Preparation Device", Chinese Patent Application No. 202520102546.1, filed on January 16, 2025, entitled "Test Tube Clamping Device and Automatic Equipment", all of which are incorporated by reference in their entirety. TECHNICAL FIELD

[0002] The present application relates to the field of biotechnology, in particular to a nucleic acid sequencing pre-treatment workstation. BACKGROUND

[0003] Nucleic acid sequencing technology is the most commonly used technology in modern molecular biology research, but complex processing procedures are required between sample extraction and machine sequencing, including nucleic acid extraction, nucleic acid extraction product quality control, library construction, library construction product quality control, and other operations. There are two traditional methods for this procedure: one is manual operation by experimenters, which has the disadvantages of high labor intensity, easy contamination, and poor consistency; the second is to use a traditional nucleic acid extraction instrument plus a single-function library construction workstation to complete, which is a semi-automatic operation, and the quality control and homogenization of the nucleic acid extraction product and the library construction product need to be completed by experimenters manually, resulting in poor user experience.

[0004] In addition, most existing nucleic acid extraction instruments are used separately. The worker places the sample in the corresponding position of the nucleic acid extraction instrument, and after the extraction is completed, the nucleic acid collected is transferred to other equipment for subsequent steps, such as a library preparation device (i.e., a library construction workstation).

[0005] In this case, the worker needs to transfer the extracted nucleic acid, which is inconvenient; if the nucleic acid extraction instrument is integrated into the library construction workstation, the aerosol generated during the nucleic acid extraction process of the sample will flow to other areas of the library construction workstation, causing contamination and affecting the quality of library construction. SUMMARY

[0006] In a first aspect, a nucleic acid sequencing pre-treatment workstation is provided, comprising:

[0007] A nucleic acid extraction system configured to controllably extract nucleic acid molecules from a biological sample;

[0008] A library construction system configured to controllably prepare the nucleic acid molecules into a library for sequencing;

[0009] A quality control system configured to controllably detect the concentration of the nucleic acid extraction product and add a normalization reagent to the nucleic acid extraction product according to the detection result and the library construction loading parameters; and / or, controllably detect the concentration of the library construction product and add a normalization reagent to the library construction product and dilute it to a concentration required for nucleic acid sequencing according to the detection result.

[0010] In a specific embodiment, the quality control system comprises a quantitative analyzer, a quality control operator assembly, a transfer mechanism, a quality control pipetting assembly, a construction reaction container, a quality control container, and a detection reagent container;

[0011] The quality control container is configured to contain the nucleic acid extraction product or the library construction product to be detected and can be placed in the quantitative analyzer to realize concentration detection;

[0012] The detection reagent container is configured to contain one or more detection reagents required for concentration detection by the quantitative analyzer and reagents for adjusting the concentration of the nucleic acid extraction product and / or the library construction product;

[0013] The transfer mechanism is configured to reciprocally transfer the quality control container between the working area of the quality control operator assembly and the working area of the quality control pipetting assembly;

[0014] The quality control operator assembly is configured to take the quality control container from the storage area of the quality control container and place it on the transfer mechanism, take the quality control container from the transfer mechanism and place it back to the storage area of the quality control container, and take the quality control container from the storage area of the quality control container and place it into the quantitative analyzer, take the quality control container that has completed detection from the quantitative analyzer and place it back to the storage area of the quality control container;

[0015] The quality control pipetting assembly is configured to suck a certain amount of detection reagent from the detection reagent container and add it to the quality control container, and suck a certain amount of reagent for adjusting the concentration of the nucleic acid extraction product and / or the library construction product and add it to the construction reaction container;

[0016] The construction reaction container is configured to contain the nucleic acid extraction product after quality control and provide a reaction space for the library construction process.

[0017] In one specific implementation, the library construction system comprises a PCR instrument, a library construction handler assembly, a library construction pipetting assembly, a library construction reagent container, a shaking and heating assembly, and a magnet assembly.

[0018] The library construction handler assembly is configured to take out the construction reaction container containing the quality-controlled nucleic acid extraction product from the quality control system, place the construction reaction container in a reagent loading area in the library construction system, and send the construction reaction container loaded with reagents to the PCR instrument; and / or send the construction reaction container containing the library construction product to the quality control system for quality control.

[0019] The library construction reagent container is configured to contain one or more library construction reagents required for library construction.

[0020] The magnet assembly is configured to provide the necessary magnetic field environment for magnetic bead purification.

[0021] The shaking and heating assembly is configured to provide a reaction space and reaction temperature for magnetic bead purification during library construction.

[0022] The library construction pipetting assembly is configured to add library construction reagents from the library construction reagent container to the construction reaction container.

[0023] In one specific implementation, it further comprises a waste container for containing the waste pipette tips of the library construction pipetting assembly.

[0024] In one specific implementation, it further comprises a rack, wherein the nucleic acid extraction system, the quality control system, and the library construction system are arranged on the workbench of the rack.

[0025] In one specific implementation, when the library construction system comprises a library construction handler assembly and a library construction pipetting assembly, the library construction handler assembly and the library construction pipetting assembly are arranged on the same three-axis motion platform, and the three-axis motion platform is arranged on the rack.

[0026] In one specific implementation, it further comprises an information acquisition module arranged on the three-axis motion platform, and the information acquisition module is used to acquire information of consumables in the nucleic acid sequencing pre-treatment workstation.

[0027] In one specific implementation, it further comprises an air filtration device arranged on the top of the rack.

[0028] In one specific implementation, the nucleic acid extraction system comprises a box with a cavity formed inside, which is composed of a bottom plate, a side plate and a top plate;

[0029] The nucleic acid extraction system further comprises a support platform and a magnetic rod sleeve assembly and a magnetic rod assembly arranged in the cavity; wherein the support platform is used to carry biological samples to be extracted and reagents required for extraction, and the magnetic rod sleeve assembly and the magnetic rod assembly are used to carry biological samples between different extraction processes;

[0030] An opening is provided on the side plate for the support platform to pass through;

[0031] Further comprising a driving member arranged on the lower side of the bottom plate, a through hole is provided on the bottom plate along a first direction, the driving member is connected with the support platform through the through hole, and the support platform is moved between the loading and unloading position and the reaction position along the first direction by the driving member;

[0032] A hatch is provided on the side plate for opening and closing the opening, and the opening is closed when the support platform is located at the loading and unloading position or the reaction position;

[0033] Wherein, the loading and unloading position is located outside the cavity, and the reaction position is located inside the cavity.

[0034] In one specific implementation, a sealing strip is provided on the bottom plate for sealing the through hole.

[0035] In one specific implementation, a rolling member is provided on the side of the support platform facing the driving member, and the support platform is connected with the driving member through the rolling member;

[0036] The sealing strip and the rolling member cooperate to block the through hole.

[0037] In one specific implementation, the rolling member comprises a support block, a connecting block and a plurality of limiting rollers;

[0038] The support block is provided with two, which is used to connect with the driving member and the support platform;

[0039] The connecting block and the limiting block are located between the two support blocks, and the two ends of the connecting block and the limiting roller are connected to the support block;

[0040] The sealing strip crosses the connecting block, and the limiting rollers are distributed on both sides of the connecting block to limit the sealing strip to cooperate with the rolling member to seal the through hole.

[0041] In one specific implementation, the driving member comprises a driving screw rod rotationally connected to the bottom plate, a sliding block sleeved on the driving screw rod, a driving motor having an output end connected to the driving screw rod, and a frame fixedly connected to the lower side of the bottom plate; the driving screw rod and the driving motor are connected to the frame.

[0042] The sliding block is threadedly engaged with the driving screw rod and fixedly connected to the object placing table.

[0043] In one specific implementation, the device further comprises a supporting frame and a plurality of consumables, the consumables being extraction operation containers.

[0044] The supporting frame is carried on the object placing table.

[0045] The plurality of consumables are arranged on the supporting frame in a first direction and detachably connected to the supporting frame.

[0046] Each of the consumables is provided with a plurality of consumable holes in a second direction, the plurality of consumable holes being respectively used for storing samples to be extracted and reagents required for extraction.

[0047] The first direction is perpendicular to the second direction.

[0048] In one specific implementation, the bottom plate is fixedly connected with a mounting rack, and the mounting rack is provided with a displacement assembly.

[0049] The displacement assembly comprises a displacement plate slidingly connected to the mounting rack in a second direction and a displacement driving member arranged on the mounting rack, the displacement driving member being connected to the displacement plate to drive the displacement plate to slide.

[0050] The magnetic rod sleeve assembly and the magnetic rod assembly are both arranged on the displacement plate.

[0051] The first direction is perpendicular to the second direction.

[0052] In one specific implementation, the magnetic rod sleeve assembly comprises a magnetic rod sleeve rack and a plurality of magnetic rod sleeves arranged on the magnetic rod sleeve rack, the magnetic rod sleeve rack being slidingly connected to the displacement plate in a third direction.

[0053] The displacement plate is further provided with a lifting driving member for driving the magnetic rod sleeve rack to slide relative to the displacement plate.

[0054] The magnetic rod assembly comprises a magnetic rod rack and a plurality of magnetic rods arranged on the magnetic rod rack, the magnetic rod rack being slidingly connected to the magnetic rod sleeve rack in the third direction.

[0055] The magnetic rod rack is provided with a secondary driving member for driving the magnetic rod rack to slide relative to the magnetic rod sleeve rack.

[0056] The third direction is perpendicular to the bottom plate, and the plurality of magnetic rods correspond to the plurality of magnetic rod sleeves one by one.

[0057] In a specific embodiment, the secondary driving member includes a screw rod threadedly engaged with the magnetic rod sleeve frame and a motor fixed relative to the magnetic rod frame, the motor being configured to drive the screw rod to rotate;

[0058] The screw rod passes through the magnetic rod frame and is rotationally connected to the magnetic rod frame, and the magnetic rod frame is fixed relative to the screw rod in the third direction.

[0059] In a specific embodiment, the system further includes a controller, a photoelectric sensor, and a warning element;

[0060] The photoelectric sensor is disposed in the box and configured to detect the position of the magnetic rod sleeve relative to the magnetic rod sleeve frame.

[0061] The controller is signal connected to the photoelectric sensor and the warning element, and is configured to trigger the warning element to issue an alarm signal when the photoelectric sensor detects that the position of the magnetic rod sleeve relative to the magnetic rod sleeve frame is deviated.

[0062] In a specific embodiment, the support frame is detachably connected to a pressing member.

[0063] The pressing member and the support frame cooperate to clamp the consumable to limit the position of the consumable.

[0064] In a specific embodiment, the consumable is provided with an identification mark for identifying a biological sample on the consumable.

[0065] In a specific embodiment, the identification mark is an identification code for scanning by an identification device, and the identification device is the information acquisition module.

[0066] In a specific embodiment, the box is provided with a plurality of ultraviolet lamps.

[0067] In a specific embodiment, the nucleic acid extraction system further includes a ventilation module, the ventilation module including an air inlet, an air outlet, and a fan disposed on the box.

[0068] The air inlet and the air outlet are located at two positions on the box that are away from each other.

[0069] The internal space of the box is in communication with the outside through the air inlet and the air outlet.

[0070] The fan is located at the air outlet, forming an air flow from the air inlet to the air outlet and flowing through the experimental area in the box.

[0071] In one specific implementation, the ventilation module further comprises a filter arranged at the air outlet for filtering the airflow.

[0072] In one specific implementation, the conveying mechanism comprises a conveying belt and at least two holding positions for holding the quality control container fixed on the conveying belt; wherein,

[0073] The conveying belt is configured to reciprocally convey the quality control container between the working area of the quality control operation hand assembly and the working area of the quality control pipetting assembly.

[0074] In one specific implementation, the quality control container is a test tube for quality control, and the holding position is a tube seat for holding the test tube.

[0075] The conveying mechanism further comprises a tube holder for elastically clamping the test tube; wherein,

[0076] The tube seat is provided with a first holding cavity with one open end; the tube holder is slidingly assembled in the first holding cavity and can slide along the axis direction of the first holding cavity.

[0077] The first holding cavity is provided with an elastic element, one end of the elastic element abuts against the tube seat, and the other end abuts against the tube holder.

[0078] The side wall of the first holding cavity is provided with an anti-rotation structure for preventing rotation of the test tube.

[0079] In one specific implementation, the tube holder has a second holding cavity for holding the test tube, the second holding cavity has one open end, and the open end of the second holding cavity is provided with a plurality of elastic clamping arms, and the plurality of elastic clamping arms are arranged at intervals around the axis of the second holding cavity.

[0080] In one specific implementation, the bottom of the second holding cavity is a conical surface.

[0081] In one specific implementation, the anti-rotation structure is a guide groove arranged on the side wall of the first holding cavity.

[0082] In one specific implementation, the first holding cavity comprises a first cavity and a second cavity in communication, and the open end of the first holding cavity is located in the first cavity; wherein, a stepped surface is formed between the first cavity and the second cavity.

[0083] The tube holder has a stepped structure limitedly matched with the stepped surface of the first holding cavity.

[0084] In an embodiment, the tube seat comprises a seat body and a support block detachably connected to the seat body.

[0085] The first cavity is located in the seat body, and the second cavity is located in the support block.

[0086] In an embodiment, the elastic element is a compression spring.

[0087] The bottom surface of the first accommodating cavity is provided with a guide column matched with one end of the compression spring.

[0088] The tube holder is provided with a ring groove matched with the other end of the compression spring.

[0089] In an embodiment, the motion mechanism is further configured to drive the tube seat to move.

[0090] In an embodiment, the motion mechanism comprises a cross beam and a slide rail arranged on the cross beam; the tube seat is slidably arranged on the slide rail; the motion mechanism further comprises a driving member configured to drive the tube seat to slide.

[0091] In an embodiment, the quality control operation hand assembly is further configured to loosen the cap of the quality control container after the quality control container is taken out from the storage area of the quality control container and placed on the conveying mechanism, or tighten the cap of the quality control container before the quality control container is taken out from the conveying mechanism and placed back to the storage area of the quality control container. BRIEF DESCRIPTION OF DRAWINGS

[0092] FIG. I is a structural block diagram of a nucleic acid sequencing pretreatment workstation provided in an embodiment of the present application;

[0093] FIG. 2 is a structural block diagram of a library construction system provided in an embodiment of the present application;

[0094] FIG. 3 is a structural block diagram of a quality control system provided in an embodiment of the present application;

[0095] FIG. 4 is a structural schematic diagram of a nucleic acid sequencing pretreatment workstation provided in an embodiment of the present application;

[0096] FIG. 5 is a schematic diagram of the arrangement of various modules of a library construction workstation provided in an embodiment of the present application;

[0097] FIG. 6 is a schematic diagram of the layout of a quality control module provided in an embodiment of the present application;

[0098] FIG. 7 is a side view of a library construction workstation provided in an embodiment of the present application;

[0099] FIG. 8 is a side view of a library construction workstation provided in an embodiment of the present application from another angle.

[0100] Fig. 9 is a schematic diagram of the overall structure of the nucleic acid extractor according to an embodiment of the present application;

[0101] Fig. 10 is a schematic diagram of the magnetic rod sleeve assembly and the magnetic rod assembly of the nucleic acid extractor according to an embodiment of the present application;

[0102] Fig. 11 is a schematic diagram of the cooperation between the sealing tape and the rolling member according to an embodiment of the present application;

[0103] Fig. 12 is a schematic diagram of the structure of the driving member and the rolling member according to an embodiment of the present application;

[0104] Fig. 13 is a schematic diagram of the structure of the support frame and the consumable according to an embodiment of the present application;

[0105] Fig. 14 is a schematic diagram of the structure of the displacement assembly driving the magnetic rod sleeve and the magnetic rod according to an embodiment of the present application;

[0106] Fig. 15 is a schematic diagram of the working principle of the photoelectric sensor according to an embodiment of the present application;

[0107] Fig. 16 is a perspective schematic diagram of the ventilation module according to an embodiment of the present application;

[0108] Fig. 17 is a schematic diagram of the cooperation between the tube seat and the tube holder according to an embodiment of the present application;

[0109] Fig. 18 is a schematic diagram of the three-dimensional structure of the tube holder according to an embodiment of the present application;

[0110] Fig. 19 is a schematic diagram of the three-dimensional structure of the test tube clamping device according to an embodiment of the present application;

[0111] Fig. 20 is a schematic diagram of the planar structure of the test tube clamping device according to an embodiment of the present application. DETAILED DESCRIPTION

[0112] In order to make the purpose, technical solutions and advantages of the present application clearer, the present application will be described in further detail below with reference to the drawings.

[0113] It should be noted that the technical terms or scientific terms used in one or more embodiments of the present specification should be understood as the general meaning understood by the person skilled in the art in the field to which the present disclosure belongs, unless otherwise defined. The terms "first", "second", and the like used in one or more embodiments of the present specification do not represent any order, quantity or importance, but are only used to distinguish different components. "Include" or "contain" and the like mean that the elements or objects before the word cover the elements or objects listed after the word and their equivalents, and do not exclude other elements or objects. "Connected" or "connected" and the like are not limited to physical or mechanical connections, but can include electrical connections, whether direct or indirect. "Up", "down", "left", "right" and the like are only used to represent relative positional relationships, and when the absolute position of the described object changes, the relative positional relationship may also change accordingly.

[0114] To facilitate the understanding of the nucleic acid sequencing pretreatment workstation provided by the embodiments of the present application, the application scenarios thereof are first described. The nucleic acid sequencing pretreatment workstation provided by the embodiments of the present application can be used in cooperation with a nucleic acid sequencer to prepare and provide a library required for nucleic acid sequencing. In the current pretreatment mode, the process required from nucleic acid extraction to library construction is relatively complex, mainly through manual operation by experimental personnel, resulting in poor consistency and stability of the pretreatment results. Therefore, the embodiments of the present application provide a nucleic acid sequencing pretreatment workstation to improve the automation of pretreatment. The following will be described in detail in combination with specific drawings and embodiments.

[0115] Referring to FIG. 1, FIG. 1 shows a structural block diagram of a nucleic acid sequencing pre-processing workstation provided by an embodiment of the present application. The nucleic acid sequencing pre-processing workstation provided by the embodiment of the present application mainly comprises a nucleic acid extraction system 1000, a library construction system 2000, and a quality control system 3000. The nucleic acid extraction system 1000 is configured to controllably extract nucleic acid molecules (such as DNA or RNA) from biological samples. The library construction system 2000 is configured to controllably prepare the nucleic acid molecules (such as DNA or RNA) into a library for sequencing. In addition, the quality control system 3000 is used to control the nucleic acid molecules. Specifically, the quality control system 3000 is configured to controllably detect the concentration of the nucleic acid extraction product, and according to the detection result and the library construction loading parameter, add a uniformization reagent to the nucleic acid extraction product; and / or, controllably detect the concentration of the library construction product, and according to the detection result, add a uniformization reagent to the library construction product and dilute it to the required concentration for nucleic acid sequencing. That is, the quality control system 3000 provided by the embodiment of the present application can only control the nucleic acid molecules extracted by the nucleic acid extraction system 1000, or only control the library constructed by the library construction system 2000, or simultaneously control the nucleic acid molecules extracted by the nucleic acid extraction system 1000 and the library constructed by the library construction system 2000.

[0116] Continuing to refer to FIG. 1, the nucleic acid sequencing pre-processing workstation provided by the embodiment of the present application further comprises an electric control system 4000 and a host computer 5000. The electric control system 4000 is configured to control the operation and work of the nucleic acid extraction system 1000, the library construction system 2000, and the quality control system 3000 according to the instructions of the host computer 5000. The host computer 5000 is configured to control the operation and work of the nucleic acid extraction system 1000, the library construction system 2000, and the quality control system 3000 by sending control instructions to the electric control system 4000. The operation and work of the above-mentioned nucleic acid extraction system 1000, library construction system 2000, and quality control system 3000 refer to the necessary actions required by the nucleic acid extraction system 1000, library construction system 2000, and quality control system 3000 to complete their corresponding functions. Through the electric control system 4000 and the host computer 5000, the control of the nucleic acid extraction system 1000, the library construction system 2000, and the quality control system 3000 is realized, so that the entire nucleic acid sequencing pre-processing workstation can be automatically operated.

[0117] As can be seen from the above description, the nucleic acid sequencing pre-processing workstation provided by the embodiment of the present application realizes the operations of nucleic acid extraction, nucleic acid extraction product quality control, library construction, and library construction product quality control through the cooperation of the nucleic acid extraction system, the quality control system, and the library construction system, improves the automation degree of the nucleic acid sequencing pre-processing process, and improves the consistency and stability of the pre-processing results.

[0118] The specific structure and work of each module in the nucleic acid sequencing pre-processing workstation provided by the embodiments of the present application will be described in detail below in combination with specific drawings and embodiments.

[0119] For the convenience of understanding, first introduce several processing processes and technical principles mainly involved in nucleic acid sequencing pre-processing.

[0120] Nucleic acid extraction is a technology for separating nucleic acid from biological samples by physical, chemical, biological methods or a combination of the above methods. It is mainly used to separate nucleic acid molecules (such as DNA or RNA) in biological samples from biological samples for subsequent experimental analysis, such as sequencing analysis.

[0121] For example, by using chemical separation, the sample cells can be lysed by a cell lysis solution to release nucleic acid molecules from the sample cells, and the nucleic acid molecules are present in the lysate solution together with impurities such as proteins. If pure nucleic acid molecules are to be obtained, further purification of the lysate is required. For example, by using magnetic bead purification, magnetic beads are added to the lysate, and the nucleic acid molecules released from the sample cells are specifically adsorbed on the surface of the magnetic beads, while the impurities such as proteins are not adsorbed and remain in the lysate. After a certain period of time, under the action of a magnetic field, the magnetic beads adsorbed with the nucleic acid molecules are separated from the solution. The magnetic beads adsorbed with the nucleic acid molecules are then washed with a washing solution to wash away the impurities such as proteins. In order to wash thoroughly, the washing can be repeated multiple times. The magnetic beads adsorbed with the pure nucleic acid molecules are recovered. Finally, the nucleic acid molecules are eluted from the magnetic beads by using an elution solution. After a certain period of time, under the action of a magnetic field, the magnetic beads are separated from the nucleic acid molecules. After removing the magnetic beads, only pure nucleic acid molecules remain in the nucleic acid extraction product.

[0122] In one implementation, for example, by using chemical separation and magnetic bead purification, the nucleic acid extraction system 1000 at least includes a nucleic acid extraction platform and a magnetic rod / magnetic rod sleeve assembly. The nucleic acid extraction platform is provided with an extraction operation container (such as a single reagent strip or a deep well plate) and a heating device (such as a heating block). The extraction operation container can contain not only one or more reagents (such as a cell lysis solution, a washing solution, and an elution solution) required for nucleic acid extraction, but also a biological sample and magnetic particles (such as magnetic beads), and provide a reaction space for various chemical reactions (such as a lysis reaction, a washing reaction, and an elution reaction) in the nucleic acid extraction process. The heating device can provide suitable reaction temperature and reaction conditions for various chemical reactions occurring in the extraction operation container. The magnetic rod sleeve is sleeved on the magnetic rod to avoid the corrosion of the magnetic rod by directly contacting the nucleic acid extraction reagent. When the magnetic rod / magnetic rod sleeve assembly enters the extraction operation container through a movement mechanism, it can provide the necessary magnetic field environment for the washing reaction and the elution reaction in the nucleic acid extraction process.

[0123] It should be noted that the nucleic acid extraction system 1000 can perform batch nucleic acid extraction processing on multiple biological samples.

[0124] When the quality control system 3000 performs quality control on the nucleic acid molecules extracted by the nucleic acid extraction system 1000, the nucleic acid extraction product containing the nucleic acid molecules can be sucked from the extraction operation container.

[0125] In the next generation sequencing (NGS) technology, the nucleic acid molecules (such as DNA or RNA) extracted from the biological sample need to be broken by physical or enzymatic methods, for example, broken by ultrasonic waves. After breaking, the nucleic acid fragments are first filled in the ends by enzymes, and then the specific enzymes are used to connect the specific DNA sequences (usually referred to as adapters) to the ends of the fragments. Finally, the nucleic acid fragments are formed and referred to as libraries in the industry.

[0126] In order to save sequencing costs, multiple biological samples will generally be sequenced simultaneously in a sequencer. In order to distinguish the sequencing results of different samples, a segment of DNA sequence (usually containing 6-8 bases) that can identify the source of the sample will be included in the adapter of the library of each sample during preparation. This segment of DNA sequence that can identify the source of the sample can also be referred to as a sample tag (or Index, Barcode). It can be understood that each sample library adapter contains its exclusive sample tag.

[0127] As shown in FIG. 2, the library construction system 2000 provided by the embodiments of the present application at least includes a PCR (Polymerase chain reaction) instrument 2100, a library construction operation hand assembly 2200 (only the working area thereof is shown in FIG. 2), a library construction pipetting assembly 2300 (only the working area thereof is shown in FIG. 2), a library construction reagent container 2400, a shaking and heating assembly 2500, and a magnet assembly 2600. It should be noted that FIG. 2 is only schematic and should not be understood as a limitation on the positions of the parts.

[0128] The library construction reagent container 2400 (e.g., a reagent kit) can contain one or more reagents required for library construction. The library construction reagent container 2400, the shaking and heating assembly 2500, and the magnet assembly 2600 are respectively located in their respective storage areas, and the respective storage areas are located within the working area of the library construction operator assembly 2200 and the library construction pipetting assembly 2300. The storage area of the library construction reagent container 2400 generally needs to be in a low-temperature refrigeration environment to facilitate low-temperature refrigeration of the library construction reagents. The shaking and heating assembly 2500 is configured to provide a reaction space and a reaction temperature for magnetic bead purification in the library construction process. The shaking and heating assembly 2500 is provided with a purification container (e.g., a deep-well plate) and a shaking and heating device. The purification container can provide a reaction space for magnetic bead purification in the library construction process, and the shaking and heating device can provide suitable reaction temperature and reaction conditions for magnetic bead purification. Meanwhile, the magnet assembly 2600 can provide a necessary magnetic field environment for magnetic bead purification. The PCR instrument 2100 is used to amplify nucleic acid molecules in the (quality-controlled) nucleic acid extraction product contained in the construction reaction container 3500.

[0129] After the nucleic acid extraction quality control is completed, the library construction operator assembly 2200 is responsible for taking the construction reaction container 3500 (to which the nucleic acid extraction product has been added) from the quality control system 3000 and placing it in a specific storage area in the library construction system 2000. In this storage area, various reaction reagents are added to the construction reaction container 3500. For the convenience of description, this storage area can be referred to as a reagent addition area. Since the construction reaction container contains nucleic acid molecules, the reagent addition area generally needs to be in a low-temperature refrigeration environment to facilitate low-temperature refrigeration of the nucleic acid molecules.

[0130] Since the entire library construction process includes multiple reaction steps, for each reaction step, the library construction pipetting assembly 2300 is responsible for first drawing the corresponding reaction reagent from the library construction reagent container 2400 and adding it to the construction reaction container 3500 (to which the quality-controlled nucleic acid extraction product has been added). The library construction operator assembly 2200 is then responsible for placing the construction reaction container 3500 (to which the nucleic acid extraction product and the corresponding reaction reagent have been added) into the PCR instrument 2100 for reaction. After the reaction is complete, the library construction operator assembly 2200 takes the construction reaction container 3500 out of the PCR instrument 2100 and places it back in the reagent addition area to wait for the addition of the next reaction reagent, and the above process is repeated.

[0131] In the library construction process, after the adapters are ligated to the ends of the nucleic acid fragments, the ligation products need to be purified to obtain a pure library. In addition, due to the small amount of library obtained at this time, library amplification is usually required. After library amplification, the amplification products also need to be purified to obtain a pure library. If magnetic bead purification is used, in addition to being able to accommodate one or more reagents required for library construction (including washing solution and elution solution required for magnetic bead purification) in the library construction reagent container 2400, magnetic beads or other magnetic particles can also be accommodated. During the magnetic bead purification reaction of the ligation product or the amplification product, the library construction pipetting assembly 2300 is responsible for first sucking the ligation product / amplification product from the construction reaction container 3500 and adding it to the purification container of the shaking and heating assembly 2500, then sucking the magnetic beads from the library construction reagent container 2400 and adding them to the purification container (to which the ligation product / amplification product has been added). After a certain period of reaction, the library construction operation hand assembly 2200 is responsible for transferring the purification container (in which the library is adsorbed on the surface of the magnetic beads) to the magnet assembly 2600. Under the action of the magnetic field, the magnetic beads adsorbed with the library are separated from the solution. The library construction pipetting assembly 2300 then sucks the supernatant in the purification container, leaving the magnetic beads adsorbed with the library. The library construction pipetting assembly 2300 continues to suck the washing solution from the library construction reagent container 2400 and add it to the purification container. After a certain period of reaction, the supernatant is again sucked away. According to experimental needs, the library construction pipetting assembly 2300 can be repeatedly operated for multiple times of washing. After washing is completed, the library construction operation hand assembly 2200 transfers the purification container to the shaking and heating assembly 2500. The library construction pipetting assembly 2300 continues to suck the elution solution from the library construction reagent container 2400 and add it to the purification container. After a certain period of reaction, the library and the magnetic beads are no longer adsorbed. The library construction pipetting assembly 2300 then sucks the supernatant (i.e., the library construction product) in the purification container and adds it to the construction reaction container 3500.

[0132] Generally, a sealing film will be attached to the upper surface of the library construction reagent container 2400 to facilitate the preservation and transportation of the internal reagents. After the reagent container is loaded into the inside of the pre-sequencing treatment instrument, the sealing film needs to be punctured by the puncturing assembly 2700 (only the working area thereof is shown in FIG. 2) to facilitate the corresponding pipetting assembly to suck the reagents from the library construction reagent container 2400. The storage area of the library construction reagent container 2400 is located within the working area of the puncturing assembly 2700.

[0133] In addition, considering that the volume of magnetic beads, washing solution, and elution solution required in the magnetic bead purification process is large and does not need to be in a low-temperature refrigeration environment, in addition to the library construction reagent container 2400, the library construction system 2000 can further be provided with a large-capacity reagent container 2900 for storing large-capacity reagents such as magnetic beads, washing solution, and elution solution.

[0134] It should be noted that the library construction operation hand assembly 2200 and the library construction pipetting assembly 2300 can be independently arranged, and the library construction operation hand assembly 2200 and the library construction pipetting assembly 2300 do not need to work in parallel, so the two assemblies can be integrated to save space. In addition, the library construction system 2000 can perform library construction processing on nucleic acid extraction products of multiple biological samples in batches.

[0135] In an implementation manner, the library construction pipetting assembly 2300 can be a pipetting pump, and a pipetting tip is provided for the pipetting pump to cooperate with the pipetting pump. When the pipetting work starts, the pipetting pump covers the pipetting tip on the end of the pipetting pump, and when the pipetting work is completed, the pipetting pump separates the pipetting tip from the end of the pipetting pump. Correspondingly, the library construction system 2000 is further provided with a pipetting tip storage area and a waste container 2800 for carrying or containing the waste pipetting tips of the library construction pipetting assembly. The used pipetting tips (waste pipetting tips) are put into the waste container 2800, as shown in FIG. 2, the waste container is arranged at the corner of the pretreatment workstation.

[0136] In an implementation manner, the library construction operation hand assembly 2200 and the library construction pipetting assembly 2300 are arranged on a three-axis motion platform, so that the library construction operation hand assembly 2200 and the library construction pipetting assembly 2300 are driven by the three-axis motion platform synchronously. For the convenience of describing the motion direction of the three-axis motion platform, an XYZ coordinate system is constructed, wherein the X direction and the Y direction and the Z direction are perpendicular to each other, and the X direction and the Y direction are parallel to the placement plane of the nucleic acid extraction system 1000, the library construction system 2000 and the quality control system 3000. The three-axis motion platform can move along the X direction, the Y direction and the Z direction respectively to meet the movement requirements of the library construction operation hand assembly 2200 and the library construction pipetting assembly 2300.

[0137] In an implementation manner, the nucleic acid sequencing pretreatment workstation provided by the embodiment of the present application further comprises an information acquisition module (for example, an industrial camera), which is arranged on the three-axis motion platform and used to acquire information of consumables in the nucleic acid sequencing pretreatment workstation. The consumables include but are not limited to extraction operation containers and library construction reagent containers. The information can be transmitted to the host computer 5000 for recording. For example, a two-dimensional code carrying consumable information is arranged on the consumable, and the industrial camera is controlled to take a photo of the two-dimensional code on the consumable, so that the host computer 5000 identifies the consumable information according to the two-dimensional code.

[0138] The concentration detection (or concentration quantitative analysis) of nucleic acid molecules (such as DNA or RNA) in the nucleic acid extraction product is important, and the total amount of nucleic acid molecules required in the subsequent library construction process is usually certain. If the actual amount of nucleic acid molecules exceeds the total amount of nucleic acid molecules required in the subsequent library construction process, the amplification efficiency during the library construction will be affected. Therefore, the concentration of nucleic acid molecules in the nucleic acid extraction product needs to be detected before the library construction. After the concentration of nucleic acid molecules is detected, the total volume of the nucleic acid extraction product required can be determined according to the concentration of nucleic acid molecules and the total amount of nucleic acid molecules required in the subsequent library construction process. Moreover, when the detected concentration of nucleic acid molecules is too high, the total volume of the nucleic acid extraction product required will be lower than the minimum volume of the pipetting device and the pipetting accuracy will be affected. Therefore, it is necessary to add a certain amount of diluent / reagent (for example, pure water) to the nucleic acid extraction product for moderate dilution before pipetting. Several common concentration detection methods include, for example, spectrophotometry, fluorescent dye detection, microfluidic analysis, or capillary gel electrophoresis.

[0139] In addition, the quality of the constructed library is crucial to the data quality of the subsequent gene sequencing output. Therefore, the quality of the library construction product needs to be detected and controlled before the sequencing. Generally, the library quality detection includes at least one of library length detection, library concentration detection, and library contaminant detection. Through the library concentration detection (or concentration quantitative analysis), the library concentration can be detected. When the detected library concentration is too high, the total volume of the library construction product required for subsequent sequencing will be lower than the minimum volume of the pipetting device and the pipetting accuracy will be affected. Therefore, it is necessary to add a diluent / reagent (for example, pure water) to the library construction product for moderate dilution before pipetting. In addition, due to the differences between the library concentrations of multiple biological samples and the differences between the sequencing data outputs, the total volume of the library construction products of multiple biological samples required for subsequent sequencing will be different and will eventually affect the balance of the sequencing data outputs of multiple biological samples. Therefore, it is also necessary to add a certain amount of diluent / reagent (for example, pure water) to the library construction product of part of the biological samples for dilution before pipetting, so as to reduce the gap between the total volumes of the library construction products of multiple biological samples. Several common concentration detection methods include, for example, spectrophotometry, fluorescent dye detection, microfluidic analysis, or capillary gel electrophoresis.

[0140] Here, it should be noted that the concentration detection of the library construction product and the nucleic acid extraction product can use the same method or different methods, which is not limited in the present application.

[0141] In the embodiments of the present application, the quality control system 3000 is used to control at least one of the library construction product and the nucleic acid extraction product. The specific structure of the quality control system 3000 will be described in detail below with reference to the drawings.

[0142] Referring to FIG. 3, FIG. 3 shows a structural schematic diagram of the quality control system provided by the embodiments of the present application. The quality control system 3000 provided by the embodiments of the present application at least includes a quantitative analyzer 3100, a quality control operator assembly 3200 (only the working area thereof is shown in FIG. 3), a conveying mechanism 3300, a quality control pipetting assembly 3400 (only the working area thereof is shown in FIG. 3), a construction reaction container 3500, a quality control container 3600, and a detection reagent container 3700. It should be noted that FIG. 3 is only schematic and should not be construed as limiting the positions of the parts.

[0143] The quality control container 3600 (for example, a centrifuge tube) can contain a detection object and can be placed in the quantitative analyzer 3100 to realize concentration detection. The detection reagent container 3700 (for example, a reagent kit) can contain one or more detection reagents required by the quantitative analyzer 3100 for concentration detection and reagents for adjusting the concentration of nucleic acid extraction products and library construction products. The construction reaction container 3500, the quality control container 3600, and the detection reagent container 3700 are respectively located in their respective storage areas. The quality control operator assembly 3200 and the quality control pipetting assembly 3400 respectively work in their respective working areas. The storage area of the quality control container 3600 is located within the working area of the quality control operator assembly 3200, and the storage areas of the construction reaction container 3500 and the detection reagent container 3700 are located within the working area of the quality control pipetting assembly 3400. The conveying mechanism 3300 is arranged between the working area of the quality control operator assembly 3200 and the working area of the quality control pipetting assembly 3400, and can reciprocally convey the quality control container 3600 between the working area of the quality control operator assembly 3200 and the working area of the quality control pipetting assembly 3400.

[0144] Before the quality control starts, the quality control operator assembly 3200 is responsible for taking the quality control container 3600 from the storage area of the quality control container 3600 and placing it on the conveying mechanism 3300. The conveying mechanism 3300 conveys the quality control container 3600 to the working area of the quality control pipetting assembly 3400. The quality control pipetting assembly 3400 is responsible for sucking a certain amount of detection reagent from the detection reagent container 3700 and adding it to the quality control container 3600. The conveying mechanism 3300 then conveys the quality control container 3600 (to which the detection reagent has been added) back to the working area of the quality control operator assembly 3200. The quality control operator assembly 3200 is then responsible for placing the quality control container 3600 (to which the detection reagent has been added) back to the storage area of the quality control container 3600.

[0145] After the start of quality control, the quality control operator assembly 3200 is responsible for taking the quality control container 3600 (which has added the detection reagent) from the storage area of the quality control container 3600 and placing it on the conveying mechanism 3300, and the conveying mechanism 3300 conveys the quality control container 3600 (which has added the detection reagent) to the working area of the quality control pipetting assembly 3400, and the quality control pipetting assembly 3400 is responsible for aspirating a certain amount of nucleic acid extraction product / library construction product from the construction reaction container 3500 and adding it to the quality control container 3600, and stirring, and then the conveying mechanism 3300 conveys the quality control container 3600 (which has added the detection reagent and the nucleic acid extraction product / library construction product in sequence) back to the working area of the quality control operator assembly 3200, and the quality control operator assembly 3200 is responsible for placing the quality control container 3600 (which has added the detection reagent and the nucleic acid extraction product / library construction product in sequence) back to the storage area of the quality control container 3600.

[0146] After the detection reagent and the nucleic acid extraction product / library construction product in the quality control container 3600 react for a period of time, the quality control operator assembly 3200 is responsible for taking the quality control container 3600 from the storage area of the quality control container 3600 and placing it into the quantitative analyzer 3100 for concentration detection, and after the detection is completed, the quality control operator assembly 3200 is responsible for taking the quality control container 3600 and placing it back to the storage area. If the detected concentration of nucleic acid molecules is higher than the required concentration, according to the detection result, the quality control pipetting assembly 3400 is also responsible for aspirating a certain amount of nucleic acid extraction product from the extraction operation container and adding it to the construction reaction container 3500, and aspirating a certain amount of pure water from the detection reagent container 3700 and adding it to the construction reaction container 3500 for dilution. Among them, the construction reaction container 3500 (such as a microwell plate) can accommodate the quality-controlled nucleic acid extraction product, and further provide a reaction space for part of the chemical reactions in the library construction process.

[0147] Similarly, if the detected library concentration is higher than the required concentration, according to the detection result, the quality control pipetting assembly 3400 is also responsible for aspirating a certain amount of pure water from the detection reagent container 3700 and adding it to the construction reaction container 3500 for dilution.

[0148] It should be noted that the quality control system 3000 can perform quality control processing on nucleic acid extraction products / library construction products of multiple biological samples in batches. In the batch processing scenario, the quality control operator assembly 3200 can take and place the quality control containers 3600 one by one, or can take and place multiple quality control containers 3600 at one time; correspondingly, the conveying mechanism 3300 can convey the quality control containers 3600 one by one, or can convey multiple quality control containers 3600 at one time; the quality control pipetting assembly 3400 can be a single-channel pipetting assembly, which sequentially aspirates and adds the detection reagent to the quality control containers 3600, or can be a multi-channel pipetting assembly, which aspirates and adds the detection reagent to multiple quality control containers 3600 at one time, and the specific configuration can be freely set according to the occupied space and work efficiency. FIG. 3 is only an example, in which the quality control operator assembly 3200 takes and places the quality control containers 3600 one by one, the conveying mechanism 3300 is provided with two container positions and can reciprocate in opposite directions to convey one quality control container 3600 each time, and the quality control pipetting assembly 3400 is a single-channel pipetting assembly.

[0149] In an implementable manner, the quality control pipetting assembly 3400 can be a pipetting pump, which is matched with the work of the pipetting pump and is also provided with a pipetting head for pipetting. When the pipetting work starts, the pipetting pump covers the pipetting head on the end thereof, and when the pipetting work is completed, the pipetting pump separates the pipetting head from the end thereof. Correspondingly, the quality control system 3000 is also provided with a pipetting head storage area, as shown in FIG. 3.

[0150] In an implementable manner, the conveying mechanism 3300 includes a conveying belt and at least two accommodation positions for accommodating the quality control containers 3600 fixed on the conveying belt; wherein the conveying belt is configured to reciprocate the accommodation positions between the working area of the quality control operator assembly and the working area of the quality control pipetting assembly to convey the quality control containers. It should be understood that the arrangement area of the conveying mechanism 3300 is not limited to the specific structure of the above example, and it can be implemented to move the quality control containers 3600 to and from the required positions.

[0151] In an implementable manner, the quality control operator assembly 3200 can be a rotating gripper, which can grasp the quality control containers 3600 and open or close the quality control containers 3600. In addition, the rotating gripper is arranged on a three-axis motion platform to move within a certain range by the three-axis motion platform to realize the necessary functions of the manipulator. The quality control pipetting assembly 3400 can also be carried on another three-axis motion platform to move to different positions by the three-axis motion platform to meet the work requirements. The movement direction of the three-axis motion platform can refer to the X direction, the Y direction and the Z direction shown in the above examples.

[0152] When the quality control system 3000 performs quality control on the plurality of quality control containers 3600, the quality control operator assembly 3200 and the quality control pipetting assembly 3400 cooperate with the transfer mechanism 3300 to open the caps of the plurality of quality control containers 3600, transfer, add liquid, and the like. For any one quality control container 3600, the quality control operator assembly 3200, the quality control pipetting assembly 3400, and the transfer mechanism 3300 disclosed in the embodiments of the present application are not sent into the quantitative analyzer 3100 after completing the addition of the quality control reagent, the addition of the nucleic acid molecule, and the mixing and stirring, and then another quality control container 3600 is operated. Instead, another quality control container 3600 is operated in the time gap of the transfer, liquid addition, or rest of the quality control container 3600, so as to improve the efficiency of the whole process.

[0153] For example, when the nucleic acid molecules are to be quality controlled, the quality control hand assembly 3200 transfers the first quality control container 3600 from the storage area to the receiving position of the conveying mechanism 3300 and rotates the cap open. The quality control hand assembly 3200 places the cap back into the first hole of the storage area. At the same time, the conveying mechanism 3300 moves the first quality control container 3600 to the working position of the quality control pipetting assembly 3400 (the position of the test reagent container 3700), and the quality control pipetting assembly 3400 loads a pipetting head in the pipetting head box, sucks the quality control reagent from the test reagent container 3700 into the quality control container 3600, and then the quality control hand assembly 3200 places the first quality control container 3600 back into the storage area. In this way, the operation of adding quality control reagents to multiple quality control containers 3600 is completed. During this process, the nucleic acid extraction system 1000 completes the extraction of nucleic acids to generate nucleic acid molecules, which are in the extraction reagent box. Then, the quality control system 3000 places the first quality control container 3600 containing the quality control reagent into the receiving position of the conveying mechanism 3300, the quality control pipetting assembly 3400 loads a pipetting head in the pipetting head box to suck the nucleic acid molecules from the elution hole of the extraction reagent box, and transfers them into the first quality control container 3600 in the corresponding receiving position of the conveying belt, and blows and mixes them. At the same time, the quality control hand assembly 3200 grabs the second quality control container 3600 from the storage area, transfers it to another receiving position of the conveying belt, and rotates the cap open. At the same time, the conveying mechanism 3300 transfers the first quality control container 3600 to the operating position of the quality control hand assembly 3200, which is screwed onto the first quality control container 3600 with the cap of the second quality control container 3600, and then transfers the first quality control container 3600 to the second hole of the storage area. After a certain period of time, the quality control hand assembly 3200 grabs the third quality control container 3600 from the storage area and places it in the receiving position of the conveying belt. During the time when the cap is grabbed and the new tube is placed, the quality control pipetting assembly 3400 loads the sample into the second quality control container 3600 and blows and mixes them. In this way, the cycle is repeated. After the first quality control container 3600 is incubated for a certain period of time, the hatch of the quantitative analyzer 3100 is opened, and the first quality control container 3600 is grabbed by the quality control hand assembly 3200 and placed into the quantitative analyzer 3100 using the interval time. The hatch of the quantitative analyzer 3100 is closed, and the host computer 5000 issues a detection instruction. After the quantitative analyzer 3100 detects, it transmits the data to the host computer 5000 for storage and processing. At the same time, the hatch of the quantitative analyzer 3100 is opened, and the first quality control container 3600 is taken out of the quantitative analyzer 3100 by the quality control hand assembly 3200 and placed back into the corresponding position of the storage area. In this way, the cycle is repeated.

[0154] In the above scheme, the transfer of each quality control container 3600 and the intervals of liquid addition and static state are utilized to operate multiple quality control containers 3600, so that the quality control system 3000 can reasonably utilize the time difference to increase the operation efficiency of the quality control containers 3600, and improve the detection efficiency of the quantitative analyzer 3100.

[0155] When the library product is subjected to quality control, the construction reaction container 3500 containing the library construction product is sent to the quality control system 3000 for quality control by the library construction operation hand assembly 2200. That is, when the library is subjected to quality control, the construction reaction container 3500 is sent to the quality control system 3000 for quality control by the library construction operation hand assembly 2200.

[0156] In an implementable scheme, the gene sequencing pre-processing workstation provided by the embodiments of the present application further includes a rack, and the nucleic acid extraction system, the quality control system, and the library construction system are arranged on a workbench 61I of the rack. The workbench 61I is used to support the nucleic acid extraction system, the quality control system, and the library construction system, so that the three systems are relatively fixed. The three-axis motion platform can be arranged on the rack to support the three-axis motion platform.

[0157] In an alternative scheme, the nucleic acid sequencing pre-processing workstation further includes an air filtration device arranged on the top of the rack. The air filtration device is installed on the top of the rack and has a fan and a high-efficiency filter. The air filtration device can filter and purify external air and then send the air into the rack to provide a clean positive pressure environment in the rack and prevent aerosol pollution. Correspondingly, air outlet fans are arranged on the left and right sides of the rack, and the air volume of the air outlet fans is less than that of the air filtration device.

[0158] To facilitate understanding of the principle of the gene sequencing pre-processing workstation provided by the embodiments of the present application, the embodiments of the present application further provide a nucleic acid sequencing pre-processing method. The method includes the following steps:

[0159] Step 001: The nucleic acid extraction system controllably extracts nucleic acid molecules from a biological sample.

[0160] For details, reference can be made to the description of the nucleic acid sequencing pre-processing workstation shown in FIGS. 1-3, which will not be described here.

[0161] Step 002: The quality control system controllably detects the concentration of the library construction product, and according to the detection result, adds a uniformization reagent to the library construction product and dilutes the library construction product to a concentration required for nucleic acid sequencing.

[0162] For details, reference can be made to the description of the nucleic acid sequencing pre-processing workstation shown in FIGS. 1-3, which will not be described here.

[0163] Step 003: The library construction system controllably prepares the nucleic acid molecules into a library for on-machine sequencing.

[0164] For details, reference can be made to the description of the nucleic acid sequencing pre-processing workstation shown in FIGS. 1-3, which will not be repeated here.

[0165] In addition, the method provided in the embodiments of the present application can further include that the quality control system controllably detects the concentration of the library in the library construction product, and adjusts the concentration of the library to be within a pre-required concentration range according to the detection result. Thus, the quality control system can control the quality of the library, so as to prepare and provide the library required for on-machine sequencing for the nucleic acid sequencer.

[0166] With continuous reference to FIGS. 4-7, FIGS. 4-7 show the structural reference diagram of the gene sequencing pre-processing workstation provided in the embodiments of the present application.

[0167] For the convenience of describing the arrangement between the modules (the nucleic acid extraction module 101 (i.e., the nucleic acid extraction system 10 in the previous embodiment), the quality detection module 201 (i.e., the quality control system 3000 in the previous embodiment), and the library construction module 401 (i.e., the library construction system 2000 in the embodiments of the present application)), an XYZ coordinate system is constructed, wherein the X direction and the Y direction and the Z direction are perpendicular to each other in pairs, and the X direction and the Y direction are both parallel to the placement plane of each module.

[0168] For the convenience of transferring the nucleic acid by the first transfer mechanism 301, when arranging the nucleic acid extraction module 101, the quality detection module 201, and the library construction module 401, the first transfer mechanism 301 is located between the nucleic acid extraction module 101, the quality detection module 201, and the library construction module 401. For example, when specifically arranging, the quality detection module 201 and the library construction module 401 are arranged side by side (along the X direction), and the nucleic acid extraction module 101 is located above the quality detection module 201 (along the Y direction). When the first transfer mechanism 301 is arranged, the first transfer mechanism 301 is located between the quality detection module 201 and the library construction module 401, and is opposite to the nucleic acid extraction module 101, that is, the quality detection module 201, the library construction module 401, and the nucleic acid extraction module 101 are arranged around the first transfer mechanism 301, so that the first transfer mechanism 301 can transfer the nucleic acid between the modules with a smaller path. The quality detection module 201 and the first transfer mechanism 301 are part of the quality control system.

[0169] With reference back to FIG. 5, the nucleic acid extraction module 101 provided by the embodiments of the present application is used to extract nucleic acid, and can be a multi-channel magnetic rod nucleic acid extraction module 101, which can simultaneously perform high-quality nucleic acid extraction on multiple samples. Specifically, the multi-channel magnetic rod nucleic acid extraction module 101 can be a magnetic rod nucleic acid extraction instrument. The nucleic acid extraction module 101 has an extraction kit 11 (i.e., a nucleic acid extraction operation container) for carrying various reagents required for nucleic acid extraction. In use, the product (containing nucleic acid) extracted by the nucleic acid extraction module 101 is carried by a test tube on the extraction kit 11. When the first transfer mechanism 301 transfers nucleic acid, the product can be sucked from the extraction kit 11 to suck the nucleic acid.

[0170] With reference to FIGS. 5 and 6, the quality inspection module 201 provided by the embodiments of the present application mainly includes a first tip box 211 (placed in a pipette head storage area), a quality inspection reagent kit 222 (i.e., a detection reagent container 3700), a quality inspection tube rack 223, a fluorescent quantifier 241 (i.e., a quantitative analyzer 3100), and a quality inspection plate site 251 (containing a reaction plate, i.e., a construction reaction container 3500); wherein the first tip box 211 is used to carry tips, which are used in cooperation with the first transfer mechanism 301 to suck nucleic acid or reagents through the tips. The quality inspection tube rack 223 is used to carry quality inspection tubes (i.e., quality control containers, which are test tubes or other tubular structures that can carry nucleic acid), which are used to carry nucleic acid and reagents. The quality inspection reagent kit 222 is used to carry quality inspection and homogenization reagents, the fluorescent quantifier 241 is used to inspect nucleic acid, and the quality inspection plate site 251 is used as a transfer structure to transfer the inspected nucleic acid to consumables “reaction plate”.

[0171] In the specific arrangement of the first tip box 21I, the quality control tube rack 23I, the quality control reagent box 22I, the quality control plate site 25I and the fluorescence quantitative instrument 24I, the first tip box 21I, the quality control reagent box 22I and the quality control plate site 25I are arranged in a single row, and specifically arranged along the X direction. In the specific arrangement, the first tip box 21I is located at the leftmost side, the quality control plate site 25I is located at the rightmost side, and the quality control reagent box 22I is located between the two. The fluorescence quantitative instrument 24I and the quality control tube rack 23I are arranged in a single row, and specifically arranged along the X direction. In the specific arrangement, the fluorescence quantitative instrument 24I is located at the left side, the quality control tube rack is located at the right side, and the fluorescence quantitative instrument 24I and the quality control tube rack 23I are located below the first tip box 21I. In the specific arrangement, the nucleic acid extraction module 10I is located above the quality control reagent box 22I and the first tip box 21I, and the first transfer mechanism 30I is arranged between the quality control reagent box 22I and the quality control plate site 25I. When arranged in the above-mentioned manner, the first transfer mechanism 30I can absorb the tip in the first tip box 21I and absorb the product in the extraction reagent box 11I in the nucleic acid extraction module 10I through a shorter path. At the same time, the quality control reagent and the uniformization reagent (such as ultrapure water) in the quality control reagent box 22I can also be absorbed through a smaller path.

[0172] With continuous reference to FIGS. 6 and 7, the first transfer mechanism 30I provided by the embodiments of the present application includes a reciprocating device 35I (i.e., a transfer mechanism 3300), a single-channel pipette 33I (i.e., a quality control pipette assembly 3400) and a rotating gripper 34I (i.e., a quality control operation hand assembly 3200). The reciprocating device 35I is used to carry the quality control tube, which includes an annular conveying belt 352I located on one side of the quality control reagent box 22I and the quality control tube rack 23I, and two hole sites 351I fixed at intervals on the annular conveying belt 352I. When carrying the quality control tube, the quality control tube can be placed on the hole site 351I. When arranging the annular conveying belt 352I, the length direction of the annular conveying belt 352I is along the Y direction, and the annular plane formed thereby is parallel to the placement plane of the nucleic acid extraction module 10I and the quality control module 20I. The annular conveying belt 352I is located between the quality control reagent box 22I and the quality control plate site 25I, and when the annular conveying belt 352I rotates, the two hole sites 351I can be moved between the quality control reagent box 22I and the quality control tube rack 23I. That is, the nucleic acid extraction module 10I, the quality control reagent box 22I, the quality control tube rack 24I and the quality control plate site 25I are all located outside the annular conveying belt 352I, so that when transplanting, the quality control tube can be transplanted through a smaller moving path.

[0173] In moving the quality inspection tube, the quality inspection tube is grabbed by rotating gripper 34I which is used to transfer the quality inspection tube between quality inspection tube rack 23I, reciprocating device 35I and fluorometric instrument 24I, and can be used to open or close the quality inspection tube. Specifically, rotating gripper 34I is used to transplant the quality inspection tube on quality inspection tube rack 23I to a hole position 351I on ring-shaped transmission belt 352I, so that the quality inspection tube can be placed on hole position 351I. In addition, rotating gripper 34I is also used to transplant the quality inspection tube after loading reagent to fluorometric instrument 24I, and transplant the quality inspection tube after completing the fluorometric detection to quality inspection tube rack 23I. To achieve the transfer of the quality inspection tube between reciprocating device 35I, fluorometric instrument 24I and quality inspection tube rack 23I.

[0174] In the process of absorbing nucleic acid or homogenizing reagent, the quality inspection tube can be opened by rotating gripper 34I, and then single-channel pipette 33I is used to achieve. Single-channel pipette 33I can be used to absorb the pipette from first pipette box 21I, and pipette in extraction reagent box 11I of nucleic acid extraction module 10I, reciprocating device 35I, quality inspection reagent box 22I and quality inspection plate position 25I. In the implementation, single-channel pipette 33I absorbs the pipette from first pipette box 21I, absorbs the extraction product in extraction reagent box 11I of nucleic acid extraction module 10I, and adds to the quality inspection tube carried in hole position 351I. In addition, it is also used to add the homogenizing reagent in quality inspection reagent box 22I to the quality inspection tube carried in hole position 351I. Then the quality inspection tube is closed by rotating gripper 34I.

[0175] It should be understood that in a specific implementation, rotating gripper 34I and single-channel pipette 33I are respectively carried on two different three-axis motion platforms. For the purpose of description, the three-axis motion platform carrying single-channel pipette 33I is named as first three-axis motion platform 31I, and the three-axis motion platform carrying rotating gripper 34I is named as second three-axis motion platform 32I. First three-axis motion platform 31I and second three-axis motion platform 32I can move in XYZ directions, wherein Z direction is perpendicular to X direction and Y direction. When rotating gripper 34I and single-channel pipette 33I are used, they can be moved to the corresponding operation position by first three-axis motion platform 31I and second three-axis motion platform 32I. For example, first three-axis motion platform 31I and second three-axis motion platform 32I can include tracks parallel to X direction, Y direction and Z direction respectively, and each track is slidingly connected with each other.

[0176] When the extraction product is transferred to the quality inspection module 20I by the first transfer mechanism 30I for inspection, the specific process is as follows: adding a quality inspection reagent to a quality inspection tube, adding the extraction product to the quality inspection tube and mixing by blowing, detecting the mixture in the quality inspection tube to obtain a quality inspection result, and then adding the extraction product and a uniformization reagent to the reaction plate according to the quality inspection result and the library construction loading parameter.

[0177] When the quality inspection reagent is added to the quality inspection tube, the first quality inspection tube is transferred from the quality inspection tube rack 23I to the hole position of the reciprocating motion device 35I by the rotating gripper 34I of the first transfer mechanism 30I, the tube cap is rotated to open, and the tube cap is placed back into the hole position of the quality inspection tube rack 23I;

[0178] The first quality inspection tube is moved to the working position of the single-channel pipettor 33I by the reciprocating motion device 35I, the single-channel pipettor 33I is used to load a suction head in a first suction head box and suck the quality inspection reagent from the quality inspection reagent box 22I to add to the first quality inspection tube, and then the first transfer mechanism 30I places the first quality inspection tube back to the quality inspection tube rack 23I, so that the reciprocating motion device 35I is used to add the quality inspection reagent to the second quality inspection tube, the third quality inspection tube and more quality inspection tubes in turn, and the first transfer mechanism 30I is used to place the second quality inspection tube, the third quality inspection tube and more quality inspection tubes back to the quality inspection tube rack 23I. At the same time in this process, the nucleic acid extraction module 10I completes the extraction of nucleic acid to generate the extraction product, and the extraction product is located in the elution hole of the extraction reagent box 11I.

[0179] Then when the extraction product is added to the quality inspection tube and mixed by blowing, the first quality inspection tube containing the quality inspection reagent is placed in the hole position of the reciprocating motion device 35I by the first transfer mechanism 30I. The single-channel pipettor 33I is used to load a suction head in a first suction head box and suck the extraction product from the corresponding elution hole of the extraction reagent box 11I to add to the first quality inspection tube and mix by blowing;

[0180] The second quality inspection tube is transferred from the quality inspection tube rack 23I to another hole position of the reciprocating motion device 35I by the rotating gripper 34I, and the tube cap is rotated to open. The first quality inspection tube is moved to the position of the rotating gripper 34I by the reciprocating motion device 35I, and the tube cap of the second quality inspection tube is covered on the first quality inspection tube. The first quality inspection tube is moved to the hole position of the quality inspection tube rack by the rotating gripper 34I.

[0181] After a first set time interval, the third quality inspection tube is grabbed from the quality inspection tube rack 23I by the rotating gripper 34I and placed in the hole position of the reciprocating motion device 35I. At the same time, the second quality inspection tube is loaded with the extraction product and mixed by blowing by the single-channel pipettor 33I.

[0182] After the first quality inspection tube is incubated for the second set time, the cabin door of the fluorescence quantitative instrument 24I is opened, the first quality inspection tube is placed into the fluorescence quantitative instrument 24I by the rotating gripper 34I at an interval, and the first quality inspection tube is taken out from the fluorescence quantitative instrument 24I by the rotating gripper 34I after detection is completed and is placed back into the corresponding position of the quality inspection tube rack 23I;

[0183] The second quality inspection tube and the third quality inspection tube are sequentially sent into the fluorescence quantitative instrument 24I for quality inspection, and are sent into the quality inspection tube rack 23I after quality inspection is completed.

[0184] As can be seen from the above description, the rotating gripper 34I, the single-channel pipettor 33I and the reciprocating device 35I cooperate to perform the work of opening the cap, transferring, adding liquid and the like on multiple quality inspection tubes. For any quality inspection tube, the rotating gripper 34I, the single-channel pipettor 33I and the reciprocating device 35I disclosed in the embodiments of the present application are not sent into the fluorescence quantitative instrument 24I after adding the quality inspection reagent, adding the extraction product, and blowing and mixing to be uniform and stationary, and then another quality inspection tube is operated, but another quality inspection tube is operated in the time interval of transferring, adding liquid or being stationary, so as to improve the efficiency of the whole process.

[0185] For example, when the extraction product is detected, the rotating gripper 34I transfers the No. 1 detection tube from the detection tube rack 23I to the hole position 351I of the reciprocating device, and rotates the cover, and the rotating gripper 34I puts the tube cover back into the No. 1 hole of the detection tube rack 23I. At the same time, the reciprocating device 35I moves the detection tube to the working position of the single-channel pipettor 33I (the position of the detection reagent kit 22I), and at the same time, the single-channel pipettor 33I loads a pipette tip in the first tip box 21I, sucks the detection reagent from the detection reagent kit 22I and adds it to the detection tube; then the first transfer mechanism 30I puts the detection tube back into the detection tube rack 23I. In this way, the operation of adding detection reagent to multiple detection tubes is completed, and in this process, the nucleic acid extraction module 10I completes the extraction of nucleic acid to generate the extraction product, which is located in the extraction reagent kit 11I. Then, the first transfer mechanism 30I places the detection tube containing the detection reagent into the hole position of the reciprocating device 35I, the single-channel pipettor 33I sucks the extraction product from the elution hole of the extraction reagent kit 11I with a pipette tip loaded in the first tip box, and transfers it to the detection tube in the corresponding hole position 351I of the reciprocating device, and blows and mixes. At the same time, the rotating gripper 34I grabs the No. 2 detection tube from the detection tube rack 23I, transfers it to the hole position 351I of the reciprocating device, and rotates the cover. Then the pipette tip is returned to the original position, and at the same time, the reciprocating device 35I transfers the detection tube to the operating position of the rotating gripper 34I, which is screwed onto the No. 1 detection tube with the tube cover of the No. 2 detection tube, and the No. 1 detection tube is transferred to the No. 2 hole of the detection tube rack 23I. After a certain period of time, the rotating gripper 34I grabs the No. 3 detection tube from the detection tube rack 23I and places it in the hole position 351I of the reciprocating device. During the time of closing the cover and grabbing a new tube, the single-channel pipettor 33I loads the sample for the No. 2 detection tube and blows and mixes, and the cycle is repeated. After the No. 1 detection tube is incubated for a proper time, the door of the fluorescence quantitative instrument 24I is opened, and the detection tube is grabbed by the rotating gripper 34I and placed in the fluorescence quantitative instrument 24I, the door of the fluorescence quantitative instrument 24I is closed, and the detection instruction is sent by the upper computer, the data is transmitted to the upper computer for saving and processing after the fluorescence quantitative instrument 24I detects, and at the same time, the door of the fluorescence quantitative instrument 24I is opened, the detection tube is taken out from the fluorescence quantitative instrument 24I by the rotating gripper 34I and placed back in the corresponding position of the detection tube rack 23I, and the cycle is repeated. In the above scheme, the operation of multiple detection tubes is performed by utilizing the transfer of each detection tube and the time interval of adding liquid and static, so that the first transfer mechanism 30I can reasonably utilize the time difference to increase the operation efficiency of the detection tube, and the detection efficiency of the fluorescence quantitative instrument 24I is improved.

[0186] After the extraction product is detected, according to the detection result and the loading parameters of the library construction, a certain proportion of nucleic acid and homogenization reagent is sucked from the elution hole of the extraction kit 11I and the detection kit 22I by the single-channel pipettor 33I and added to the consumables "reaction plate" on the detection plate site 25I. The product detection process after library construction is the same as above, except that the sample position is changed from the extraction kit 11I to the detection plate site 25I; during the homogenization operation, according to the concentration detection data in the upper computer, the homogenization reagent in the detection kit 22I is sucked by the single-channel pipettor 33I and added to the consumables "reaction plate" on the detection plate site 25I, and the library construction product is diluted to the required concentration for sequencing.

[0187] In an implementable scheme, the two hole sites 351I are arranged at intervals. The arrangement position of the two hole sites 351I satisfies: when the detection tube carried by one of the hole sites 351I is loaded with nucleic acid, the other hole site 351I can be moved to the working area of the rotary clamp jaw 34I, so that the movement of the rotary clamp jaw 34I and the single-channel pipettor 33I does not interfere with each other, so as to make full use of the time of the single-channel pipettor 33I to load the sample and mix evenly by blowing, and to carry out the operations such as opening and closing the cap, transferring the detection tube and the like by the rotary clamp jaw 34I, thereby improving the detection efficiency.

[0188] Referring to FIG. 5 and FIG. 8, the library construction module is used to construct a library of nucleic acid. The library construction module 40I mainly includes a second tip box 41I (placed in the pipette tip storage area), a reagent box 42I (i.e., a large volume reagent container 2900), a shaking module 43I (i.e., a shaking and heating assembly 2500), a sample loading plate 45I (i.e., a reagent loading area), a magnetic stand 46I (i.e., a magnet assembly 2600), a refrigeration module 48I (accommodating a library construction reagent container 2400), and a second transfer mechanism 50I. The second tip box 41I is used to carry the tips; the reagent box 42I is used to carry the large volume reagents required for library construction, the sample loading plate 45I is used to maintain a low temperature environment when adding reagents in the consumables “reaction plate” to prevent premature reaction of the reagents, and the refrigeration module 48I can refrigerate and store the key reagents required for library construction to prevent deterioration; the magnetic stand 46I and the shaking module 43I are used for magnetic bead purification of nucleic acid to meet the efficient magnetic bead purification in the library construction process. Specifically, the reagent box 42I, the magnetic stand 46I, and the shaking module 43I are used for magnetic bead purification in the library construction process. The second transfer mechanism 50I is used to flexibly transfer the consumables (which at least include the reaction plate and the purification plate) among the quality inspection plate 25I (accommodating the reaction plate, which is the construction reaction container 3500), the sample loading plate 45I, the thermal cycling module 47I (i.e., a PCR instrument 2100), the magnetic stand 46I, and the shaking module 43I, and is used to load the reagents in the refrigeration module 48I and the reagent box 42I into the consumables “reaction plate” and “purification plate” (i.e., purification container). The functions of the above-mentioned second tip box 41I, reagent box 42I, refrigeration module 48I, magnetic stand 46I, and shaking module 43I are all common modules in the library construction module, which will not be described in detail in the embodiments of the present application.

[0189] In addition, the library construction module 40I provided by the embodiments of the present application can also include a thermal cycling module 47I, which is fixed on the workbench 61I and is used to provide rapid and high-precision temperature control for the library construction module. In a specific arrangement, the thermal cycling module 47I is also located at the corner of the workbench 61I, and an air outlet fan 30I1 is arranged on the right side of the thermal cycling module 47I, which can quickly extract the generated aerosol to reduce the risk of pollution.

[0190] In addition, the library construction module 40I also includes a waste box 44I (i.e., a waste container 2800), which is used to hold the waste generated in the library construction process. In a specific arrangement, the waste box 44I is diagonally arranged with the nucleic acid extraction module 10I, so as to reasonably utilize the space. In addition, the waste box 44I is located at the corner position, which is relatively far away from other modules. This reduces the probability of waste contaminating other substances.

[0191] With reference to FIGS. 5, 7 and 8, when arranging the various plate positions in the library construction module 40I, the second tip box 41I, the reagent box 42I, the shaking module 43I, the waste box 44I, the sample addition plate position 45I, the magnetic stand 46I, the thermal cycling module 47I, and the cold storage module 48I are arranged in an array. As shown in FIG. 5, the second tip box 41I, the reagent box 42I, the shaking module 43I, the waste box 44I, the sample addition plate position 45I, the magnetic stand 46I, the thermal cycling module 47I, and the cold storage module 48I are arranged in an array, and the whole forms a stepped structure to avoid the structures in the nucleic acid extraction module 10I and the quality inspection module 21I, so that the various plate positions of the workstation form an arrayed whole structure on the placement surface.

[0192] In a specific arrangement, the second tip box 41I is arranged in multiple rows and in a stepped manner. The second tip box 41I forms a stepped structure to avoid the nucleic acid extraction module 10I, the quality inspection module 20I, and the cold storage module 44I. As shown in FIG. 5, the second tip box 41I is arranged in three rows, and each row is arranged along the Y direction. The multiple rows of second tip boxes 41I are arranged along the X direction. Along the X direction, there is one second tip box 41I in the first row, five second tip boxes 41I in the second row, and three second tip boxes 41I in the third row, thereby forming a stepped arrangement. When matched with the quality inspection module 20I and the nucleic acid extraction module 10I, the second tip box 41I in the first row is arranged in a row along the Y direction with the quality inspection plate position 25I. The second tip boxes 41I in the second and third rows are located on one side of the nucleic acid extraction module 10I, thereby reasonably utilizing the space to integrate the nucleic acid extraction module 10I, the quality inspection module 20I, and the library construction module 40I on a placement surface, and to arrange the three modules closely.

[0193] The cold storage module 48I has multiple, as shown in FIG. 5, the number of cold storage modules 42I is 4, and the 4 cold storage modules 48I are arranged in two rows along the Y direction, two by two in a row, which are located on the right side of the second row of second tip boxes 41I, above the third row of second tip boxes 41I and the sample addition plate position 45I.

[0194] The sample addition plate position 45I, the reagent box 42I, and the shaking module 43I are arranged in a row along the Y direction, and in terms of sequence, from top to bottom, they are the sample addition plate position 45I, the reagent box 42I, and the shaking module 43I. The thermal cycling module 47I and the magnetic stand 46I are arranged in a row along the Y direction, and in terms of sequence, from top to bottom, they are the thermal cycling module 47I and the magnetic stand 46I.

[0195] During the library construction process, the consumables "reaction plate" and "purification plate" can be transferred between the quality inspection plate site 25I, the sample adding plate site 45I, the thermal cycle module 47I, the magnetic stand 46I and the oscillation module 43I by the second transfer mechanism 50I, and the reagents in the refrigeration module 48I and the reagent box 43I can be loaded into the consumables "reaction plate" and "purification plate".

[0196] In a specific arrangement, the second transfer mechanism 50I includes a multi-channel pipettor 52I, which is used to suck the pipette tips in the second tip box 41I and to transfer liquid in each plate site of the library construction module.

[0197] In a specific use, the second transfer mechanism 50I includes a three-axis motion platform, which is named as the third three-axis motion platform 51I for convenience of description. The third three-axis motion platform 51I can realize movement in the X direction, the Y direction and the Z direction. In addition, the second transfer mechanism 50I further includes a transfer plate clamping jaw 53I (i.e., the library construction operation hand assembly 2200) arranged on the third three-axis motion platform 51I; and the multi-channel pipettor 52I (i.e., the library construction pipetting assembly 2300) is also arranged on the third three-axis motion platform 51I. The transfer plate clamping jaw 53I can flexibly transfer the consumables "reaction plate" and "purification plate" between the quality inspection plate site 25I, the sample adding plate site 45I, the thermal cycle module 47I, the magnetic stand 46I and the oscillation module 43I.

[0198] In an optional solution, the library construction module further includes an information acquisition module 54I arranged on the multi-channel pipettor 52I, which is used to acquire the information of the reagent box when the pipettor collects the reagent. The information is uploaded to the upper computer to realize information acquisition. In addition, the information acquisition module 54I can also acquire the information of the consumables and the position information of the motion mechanism (the second transfer mechanism 50I). The upper computer judges whether the consumables are misplaced or missed according to the information acquired by the information acquisition module 54I, and calibrates the position deviation of the motion mechanism according to the information acquired by the information acquisition module 54I. It should be understood that the execution of the upper computer according to the information acquired by the information acquisition module 54I is a conventional control mode in the art, which will not be described here.

[0199] In order to carry the above-mentioned modules, the workstation provided by the embodiment of the application further includes a rack 60I, and the rack 60I is provided with a workbench 61I. The nucleic acid extraction module 10I, the quality inspection module 20I and the library construction module 40I are arranged on the workbench 61I. The workbench 61I carries the above-mentioned modules. In addition, the three-axis motion platforms in the first transfer mechanism 30I and the second transfer mechanism 50I are supported by the rack 60I, and the transfer plate clamping jaw 53I and the rotary clamping jaw 33I are suspended above the workbench 61I, so that the clamped quality inspection tube can be moved between different modules.

[0200] The rack 601 is also provided with a sealed cabin door 621, and the sealed cabin door 621 and the rack instrument isolate the air in the instrument from the outside of the instrument, thereby preventing sample contamination caused by the external environment, and also preventing nucleic acid contamination in the instrument from the external environment.

[0201] In an alternative embodiment, the workstation further comprises an air filtration device 641 provided on the top of the rack 601. The air filtration device 641 is installed on the top of the rack 601 and has a fan and a high-efficiency filter, which can filter and purify the external air and then send it into the cabinet to provide a clean positive pressure environment in the cabinet, thereby preventing aerosol contamination. Correspondingly, the left and right sides of the rack 601 are provided with air outlet fans 651, and the air volume of the air outlet fans 651 is less than that of the air filtration device 641.

[0202] In order to facilitate the understanding of the workstation provided in the embodiments of the present application, the embodiments of the present application also provide a method for constructing a nucleic acid library, which utilizes any of the workstations described above; the method comprises the following steps:

[0203] Step 001: performing nucleic acid extraction by the nucleic acid extraction module and forming an extraction product;

[0204] Specifically, reference is made to the related descriptions in FIGS. 4-8, which are not repeated here.

[0205] Step 002: transferring the extraction product to the quality inspection module by the first transfer mechanism for quality inspection;

[0206] Specifically, the following steps are included:

[0207] Step a: adding a quality inspection reagent to the quality inspection tube;

[0208] Specifically, the quality inspection tube is transferred from the quality inspection tube rack to the hole site of the reciprocating motion device by the rotating clamping jaw of the first transfer mechanism, and the tube cap is opened by rotation;

[0209] The quality inspection tube is moved to the working position of the single-channel pipettor by the reciprocating motion device; the single-channel pipettor is used to load a pipette tip in the first tip box and to suck the quality inspection reagent from the quality inspection reagent box and add it to the quality inspection tube; then the first transfer mechanism returns the quality inspection tube to the quality inspection tube rack;

[0210] The above steps are repeated until the quality inspection reagent is added to a predetermined number of quality inspection tubes.

[0211] Step b: adding the extraction product to the quality inspection tube and mixing by blowing;

[0212] Specifically, the first quality inspection tube is placed into the hole position of the reciprocating device by the first transfer mechanism, the tube cap is rotated open, and the tube cap is placed back into the hole position of the quality inspection tube rack; the single-channel pipettor loads the pipette tips in the first tip box and sucks the extraction product from the corresponding elution hole of the extraction kit and adds it into the first quality inspection tube for blowing and mixing;

[0213] The second quality inspection tube is transferred from the quality inspection tube rack to another hole position of the reciprocating device by rotating the clamping jaw, and the tube cap is rotated open; the first quality inspection tube is moved to the position of the rotating clamping jaw by the reciprocating device, and the tube cap of the second quality inspection tube is closed on the first quality inspection tube; and the first quality inspection tube is moved to the hole position of the quality inspection tube rack by the rotating clamping jaw;

[0214] After a first set time interval, the third quality inspection tube is grabbed from the quality inspection tube rack by the rotating clamping jaw and placed in the hole position of the reciprocating device; at the same time, the single-channel pipettor sucks the extraction product from the corresponding elution hole of the extraction kit and adds it into the second quality inspection tube for blowing and mixing;

[0215] The above steps are repeated until a predetermined number of quality inspection tubes are added with the extraction product of the corresponding elution hole and are blown and mixed.

[0216] Step c, detecting the mixture of the quality inspection tube to obtain a quality inspection result;

[0217] Specifically, after the first quality inspection tube is incubated for a second set time, the cabin door of the fluorescence quantitative instrument is opened, the rotating clamping jaw moves the first quality inspection tube and places it into the fluorescence quantitative instrument for detection, and after the detection is completed, the first quality inspection tube is taken out from the fluorescence quantitative instrument by the rotating clamping jaw and placed back into the corresponding position of the quality inspection tube rack;

[0218] The above steps are repeated until the quality inspection of the predetermined number of quality inspection tubes is completed.

[0219] Step d, adding the extraction product and the uniformization reagent into the reaction plate according to the quality inspection result and the library construction loading parameter.

[0220] According to the quality inspection result of each quality inspection tube and the library construction loading parameter, the single-channel pipettor sucks a certain proportion of the extraction product and the uniformization reagent from the corresponding elution hole of the extraction kit and the quality inspection kit, respectively, and adds them into the corresponding hole position of the reaction plate on the quality inspection plate.

[0221] Specifically, refer to the related description in FIGS. 4-8, which will not be repeated here.

[0222] Step 003: transferring the nucleic acid after the quality inspection of the quality inspection module to the library construction module for library construction by the second transfer mechanism.

[0223] Specifically, refer to the relevant description in FIGS. 4-8, which will not be repeated here.

[0224] In the above technical solution, through the cooperation of the nucleic acid extraction module, the quality inspection module, the library construction module, and the first transfer mechanism, nucleic acid extraction, quality inspection, library construction, quality inspection, and uniformization operations are realized, improving the automation level of library construction, thereby improving the consistency of the library and reducing the influence of external risks on the detection results.

[0225] To facilitate understanding of the nucleic acid extractor provided in the embodiments of the present application, the application scenario thereof is first described. The nucleic acid extractor provided in the embodiments of the present application is applied to the extraction of nucleic acid. Most of the existing nucleic acid extractors are used independently. The staff places the sample in the corresponding position of the nucleic acid extractor, and after the extraction is completed, the extracted nucleic acid is collected and transferred to other equipment for subsequent other steps, such as a library preparation device (i.e., a library construction workstation). In this case, the staff needs to transfer the extracted nucleic acid, which is relatively inconvenient. If the nucleic acid extractor is integrated into the library preparation device (forming an all-in-one machine or a pre-treatment platform), the aerosol generated during the nucleic acid extraction process of the sample will flow to other areas of the library preparation device, causing pollution and affecting the quality of library construction. Therefore, the embodiments of the present application provide a nucleic acid extractor, a library preparation device, and a nucleic acid extraction process to overcome the problem of resource waste. The specific drawings and embodiments will be described in detail below.

[0226] First, refer to FIGS. 9 and 10, which show the main structural schematic diagram of the nucleic acid extractor (i.e., the nucleic acid extraction system in the all-in-one machine), which is used for extracting nucleic acid from a sample to obtain target nucleic acid. The nucleic acid extractor includes a box body 1 with a cavity formed inside, which is composed of a bottom plate 11 and side plates and a top plate. One of the side plates is provided with an opening and a hatch 12 for opening and closing the opening to realize the taking and placing operations of samples, reagents, and other items. The bottom plate 11 is provided with a placement table 2, a magnetic bar sleeve assembly 3, and a magnetic bar assembly 4. The placement table 2 is used to carry the sample to be extracted, and the magnetic bar sleeve assembly 3 and the magnetic bar assembly 4 are used to transfer the sample carried by the magnetic beads between different extraction processes.

[0227] Referring to FIGS. 9 and 10, the placement table 2 is slidably connected with the bottom plate 11 along the first direction and can be stopped at a corresponding position relative to the bottom plate 11; in addition, the nucleic acid extractor further comprises a driving member 21 arranged on the lower side of the bottom plate 11 and used to drive the movement of the placement table 2, the driving member 21 drives the placement table 2 to pass through the opening on the box body 1 and slide in the direction inside and outside the box body 1. When the sample is placed on the placement table 2 and the subsequent extraction work is completed and the target nucleic acid is taken away, the placement table 2 is located outside the box body 1, which reduces the influence of factors such as narrow space and makes the taking and placing operation more convenient.

[0228] The bottom plate 11 is provided with a communication hole extending along the first direction, the placement table 2 is located on the upper side of the bottom plate 11, the driving member 21 is located on the lower side of the bottom plate 11, the driving end of the driving member 21 is connected with the placement table 2 through the communication hole, the driving of the placement table 2 is realized, and the placement table 2 is driven to move between the taking and placing position and the reaction position in the first direction. The taking and placing position is located outside the cavity of the box body 1 and is used to take and place the articles on the placement table 2; the reaction position is located inside the cavity of the box body 1 and is used to perform the extraction process on the sample.

[0229] Exemplarily, referring to FIGS. 11 and 12, the driving member 21 is a screw block structure, which comprises a driving motor 21a, a driving screw 21b and a sliding block 21c. The driving screw 21b extends along the first direction and is rotationally connected to the lower side of the bottom plate 11. The sliding block 21c is provided with a threaded hole through which the driving screw 21b passes. The driving screw 21b is arranged in the threaded hole and is in threaded connection with the sliding block 21c. In addition, the sliding block 21c is fixedly connected with the placement table 2 through the communication hole. The driving motor 21a is fixedly connected relative to the bottom plate 11. The driving motor 21a is connected with the driving screw 21b to drive the rotation of the driving screw 21b.

[0230] The bottom plate 11 is fixedly connected with a frame 11a on the lower side, the driving motor 21a is fixedly connected in the frame 11a, and the driving screw 21b is rotationally connected with the frame 11a. In addition, the frame 11a is further provided with a limiting block 11b which is attached to the inner side wall of the frame 11a. The sliding block 21c is fixedly connected with the limiting block 11b, which can improve the stability of the sliding block 21c under the driving of the driving screw 21b.

[0231] In this way, the driving member 21 can be conveniently arranged to adjust the position of the placement table 2. In addition, the driving member 21 is arranged on the lower side of the bottom plate 11, which reduces the occupation of the internal space of the box body 1 and improves the utilization rate of the internal space.

[0232] In addition, the driving element 21 is arranged on the lower side of the bottom plate 11, which does not occupy the space of the internal cavity of the box body 1, and when the driving element 21 drives the placement platform 2 to move to the taking and placing position or the reaction position, the opening on the box body 1 can be closed by using the hatch 12 to isolate the internal cavity of the box body 1 from the external environment, reduce the diffusion of aerosol generated in the reaction process to the surrounding environment, and reduce the pollution problem caused to the surrounding environment. Therefore, when the nucleic acid extractor is integrated in the library preparation device, it is not easy to cause pollution to other areas, which is beneficial to improve the quality of library construction and reduce the problem that the nucleic acid extractor and the library preparation device cannot be integrated due to pollution.

[0233] In order to ensure that the cavity in the box body 1 has better sealing performance, the bottom plate 11 is provided with a sealing strip 13 for sealing the communication hole.

[0234] By arranging the sealing strip 13, the cavity in the box body 1 has better sealing effect, which avoids the communication hole always communicating with the outside, so that the space in the box body 1 is easily polluted, and avoids the pollution of the finally extracted nucleic acid, and the extraction effect is better.

[0235] Specifically, the side of the placement platform 2 facing the driving element 21 is provided with a rolling element 22, and the placement platform 2 is connected with the driving element 21 through the rolling element 22; and the sealing strip 13 cooperates with the rolling element 22 to block the communication hole, which improves the sealing performance of the box body 1 and further reduces the pollution problem caused by the diffusion of aerosol to the outside.

[0236] Referring to FIG. 12, the rolling element 22 includes a support block 22a, a connecting block 22b and a limiting roller 22c, wherein the support block 22a is provided with two and is fixedly connected with the placement platform 2, the connecting block 22b and the limiting roller 22c are located between the two support blocks 22b, and the two ends of the connecting block 22b and the limiting roller 22c are connected with the two support blocks 22b, so that the rolling element 22 forms an integral whole, and the connecting block 22b and the limiting roller 22c are spaced apart from the placement platform 2. The sealing strip 13 crosses the connecting block 22b and is limited in position by the limiting roller 22c, so that the sealing strip 13 cooperates with the rolling element 22 to seal the communication hole.

[0237] For example, the limiting roller 22c has two and is rotatably connected with the support block 22a, and the two limiting rollers 22c are located on both sides of the connecting block 22b in the extension direction of the sealing strip 12; the sealing strip 13 extends from the lower side of one of the limiting rollers 22c to the upper side of the connecting block 22b, and passes through the lower side of the other limiting roller 22c. By limiting the sealing strip 13 with the limiting roller 22c, the sealing performance of the sealing strip 13 to the communication hole can be guaranteed.

[0238] In addition, the edge of the connecting block 22b that is connected with the sealing band 13 is provided with an arc transition, the sealing band 13 crosses the connecting block 22b and is attached to the arc on the upper side of the connecting block 22b, so that the sealing band 13 slides more smoothly relative to the connecting block 22b during the sliding of the placement table 2, reducing the wear on the sealing band 13.

[0239] The rolling part 22 is arranged to stably position the sealing band 13, and during the sliding of the placement table 2, the sealing band 13 is always in cooperation with the rolling part 22 to seal the communication hole, and the sealing effect of the whole box 1 is better.

[0240] Referring to FIG. 9, the bottom plate 11 has an extension 14 on the side of the opening, which extends away from the cavity in the box 1, so that the placement table 2 can slide to the outside of the box 1, and when the placement table 2 slides onto the extension 14, it is in the taking and placing position.

[0241] In this way, in addition to closing the hatch 12 during nucleic acid extraction to isolate the space in the box 1 from the outside world and avoid contamination of the outside world during nucleic acid extraction, the hatch 12 can also be closed before and after the extraction operation, and then the sample, nucleic acid, etc. placed on the placement table 2 can be taken and placed, reducing the pollution of the reagents in the nucleic acid extractor to the external environment and reagents.

[0242] Referring to FIG. 9, the hatch 12 is sealingly connected with the box 1, and the box 1 and the hatch 12 are provided with an opening and closing driving part 15 for driving the hatch 12 to move relative to the box 1 to open and close the opening in the box 1. The opening and closing driving part 15 can be a linear driving structure such as a pneumatic cylinder, a hydraulic cylinder, an electric push rod, or a transmission structure such as a lead screw and a sliding block, a gear and a rack, a chain, a worm and a gear. Only part of the structure of the opening and closing driving part 15 is shown in FIG. 9, and the actual arrangement can be designed according to the existing structure in the related field, as long as it can achieve the opening and closing of the hatch 12.

[0243] Further improves the automation degree of the nucleic acid extractor, and is more convenient to use. In addition, through this arrangement, the nucleic acid extractor can be integrated with other equipment.

[0244] For example, this nucleic acid extractor can be integrated into other nucleic acid testing equipment (i.e., a library construction workstation). During the process, the operator only needs to manually place the sample on the corresponding consumable 6. After that, no further manual operation is required. The stage 2 is automatically moved to the corresponding position inside the housing 1, and the door 12 is closed for extraction. After extraction, the door 12 automatically opens, moving the stage 2 out of the housing 1. A robotic arm then transfers the extracted target nucleic acid to the subsequent testing process. In this process, except for sample placement, the remaining steps are all automated, greatly improving ease of use. Furthermore, since no manual operation is required, the corresponding steps are completed in an environment isolated from the outside world, further reducing the risk of contamination of samples and target nucleic acids, resulting in better extraction results.

[0245] Referring to Figure 13, the nucleic acid extractor also includes a support frame 5 and multiple consumables 6 (the consumables 6 are extraction operation containers, such as single reagent strips or deep well plates). The support frame 5 is supported on the stage 2. For example, the support frame 5 can be placed directly on the stage 2, or the position of the support frame 5 relative to the stage 2 can be limited by structures such as buckles, slots, and limiting plates.

[0246] The support frame 5 is provided with placement holes for accommodating consumables 6. In actual installation, the number of placement holes on a support frame 5 can be adjusted according to actual needs. Referring to Figure 13, the support frame 5 is provided with four placement holes, and each placement hole can accommodate a set (or more) of consumables 6.

[0247] The main body of the consumable 6 is located in the placement hole. The two ends of the consumable 6 that are far apart from each other are integrally formed with overlapping edges. The overlapping edges are attached to the support frame 5. Specifically, the overlapping edges are attached to the edge of the placement hole.

[0248] In one embodiment, a clamping member 51 is detachably connected to the support frame 5. The clamping member cooperates with the support frame to clamp the overlapping edge of the consumable 6 to restrict the position of the consumable. The clamping member 51 and the support frame 5 can be connected by means of snaps, magnets, etc. In actual use, the consumable 6 may have a sealing film affixed to it to seal each consumable hole 61. By setting the clamping member 51, the position of the consumable 6 relative to the support frame 5 is restricted. In this way, it is avoided that when the sealing film on the consumable 6 is peeled off, the consumable 6 will be lifted off the support frame 5, making the operation more convenient. It can also prevent problems such as the internal sample splashing out due to the consumable 6 being lifted and then falling.

[0249] For example, a limiting part is provided on the inner side of the placement hole. The position of each consumable is determined by the limiting part, which can ensure the positional accuracy of the consumable 6 relative to the support 5 and avoid positional deviation that would interfere with the subsequent extraction process.

[0250] The consumables 6 are provided with a plurality of consumable holes 61 in the second direction, respectively used for storing samples to be extracted and various reagents required for extraction. Exemplarily, the plurality of consumable holes 61 are divided into sample slots, magnetic bead slots, lysis slots, rinsing slots and elution slots, wherein the number of rinsing slots can be set as needed to perform multiple rinsing processes during extraction, which is beneficial to improve the purity of the target nucleic acid obtained by elution.

[0251] For the convenience of subsequent description, the following is explained: the first direction is the moving direction when the article is put into or taken out of the box 1 through the opening on the box 1, which is in the horizontal plane; the second direction is also in the horizontal plane and perpendicular to the first direction. In addition, a third direction is also set, which is perpendicular to the plane in which the first direction and the second direction lie, i.e. the up-down direction in the reagent use state of the nucleic acid extractor.

[0252] In actual use, according to the number of samples to be extracted, the corresponding number of consumables 6 are installed on the support frame 5, and samples, magnetic beads or various corresponding reagents are placed in different consumable holes 61. Then, the support frame 5 with consumables 6 is placed on the workbench 2, and the subsequent extraction process is performed. As can be seen, when the nucleic acid extractor is used for nucleic acid extraction, the corresponding number of consumables 6 can be used according to the number of samples to be extracted, and there is no limitation that only 16 (or 8, or other specified number) samples can be extracted at a time. The nucleic acid extractor can be used to extract one, two, three or other different number of samples conveniently. Moreover, when different number of samples are extracted, there is no pollution of reagents in other holes corresponding to the samples to be extracted, which reduces the problem of reagent waste.

[0253] The consumables 6 are provided with identification marks for identifying the samples on the corresponding consumables 6. In actual use, the identification marks are set to facilitate the differentiation of different samples. Exemplarily, the identification marks are identification codes for scanning by an identification device (i.e. an information acquisition module), such as bar codes, two-dimensional codes, etc. When nucleic acid extraction is performed, the identification codes on the consumables 6 are scanned, which can conveniently distinguish different consumables 6 (samples).

[0254] In addition, a plurality of ultraviolet lamps are arranged in the box 1, which can disinfect the space in the box 1 before and after the experiment, prevent pollution, improve the purity of the extracted nucleic acid, and reduce the risk of low experimental accuracy caused by pollution.

[0255] Referring to FIG. 14, the mounting rack 16 is fixedly connected to the inner bottom plate 11 of the box 1, the displacement assembly 7 is arranged on the mounting rack 16, the magnetic rod sleeve assembly 3 and the magnetic rod assembly 4 are both mounted on the displacement assembly 7, and the magnetic rod sleeve assembly 3 and the magnetic rod assembly 4 are driven to move in the second direction by the displacement assembly 7. The displacement assembly 7 comprises a displacement plate 71 which is slidably connected to the mounting rack 16 in the second direction and a displacement driving member 72 which is fixedly connected to the mounting rack 16, the displacement driving member 72 is connected to the displacement plate 71 to drive the displacement plate 71 to slide in the second direction, and the magnetic rod sleeve assembly 3 and the magnetic rod assembly 4 are arranged on the displacement plate 71. Exemplarily, the displacement driving member 72 is a screw block structure, which can drive the displacement plate 71 to slide relative to the mounting rack 16 in the same manner as the driving member 21 drives the placement table 2 to move, and the structure thereof will not be described in detail herein.

[0256] The displacement assembly 7 drives the magnetic rod sleeve assembly 3 and the magnetic rod assembly 4 to move in the second direction, so as to correspond to different consumable holes 61 on the consumable 6 and drive the magnetic beads to transfer between the different consumable holes 61.

[0257] Referring to FIG. 14, the magnetic rod sleeve assembly 3 comprises a magnetic rod sleeve rack 31 and a plurality of magnetic rod sleeves 32 mounted on the magnetic rod sleeve rack 31, and the mounting here refers to that the magnetic rod sleeves 32 are detachably and fixedly connected to the magnetic rod sleeve rack 31. The magnetic rod sleeve rack 31 is slidably connected to the displacement plate 71 in the third direction, and a lifting driving member 74 is fixedly connected to the displacement plate 71, and the lifting driving member 74 is connected to the magnetic rod sleeve rack 31 to drive the magnetic rod sleeve rack 31 to slide relative to the displacement plate 71.

[0258] Exemplarily, the lifting driving member 74 comprises a lifting plate which is slidably connected to the displacement plate 71 in the third direction, and the magnetic rod sleeve assembly 3 is fixed to the lifting plate. The lifting driving member 74 drives the lifting plate to slide in the form of belt driving, specifically, the lifting driving member 74 further comprises two pulleys which are rotatably connected to the displacement plate 71, a belt which is tautly arranged on the two pulleys, and a motor which is fixedly connected to the displacement plate 71. One side of the lifting plate is fixedly connected to the belt, the motor drives the belt to rotate, and drives the lifting plate to slide and lift in the third direction. In actual use, in order to ensure the stability of driving the magnetic rod sleeve rack 31 to slide, the belt is a synchronous belt, that is, the inner side of the belt and the pulleys are provided with toothed structures which cooperate with each other.

[0259] The magnetic rod assembly 4 comprises a magnetic rod rack 41, a plurality of magnetic rods 42 mounted on the magnetic rod rack 41, and a secondary driving member 43 arranged on the lifting plate. The secondary driving member 43 is used to keep the magnetic rod rack 41 stationary relative to the lifting plate and the magnetic rod sleeve rack 31, or drive the magnetic rod rack 41 and the magnetic rods 42 to slide relative to the lifting plate and the magnetic rod sleeve 32. The magnetic rod sleeve 32 is used to extend into different placement holes, and the plurality of magnetic rods 42 correspond to the plurality of magnetic rod sleeves 32 one by one.

[0260] The secondary driving member 43 comprises a screw rod threadedly matched with the magnetic rod sleeve frame 31 and a motor fixedly connected to the lifting plate, and the motor is used to drive the screw rod to rotate; the screw rod penetrates through the magnetic rod frame 41 and is rotationally connected with the magnetic rod frame 41, and the magnetic rod frame 41 is relatively fixed with the screw rod in the third direction. Here, the magnetic rod frame 41 is relatively fixed with the screw rod in the third direction, which means that the two only rotate relative to each other, but do not slide relative to each other in the third direction. In this way, the motor drives the screw rod to rotate, and drives the magnetic rod frame 41 to slide relative to the magnetic rod sleeve frame 31 in the third direction, thereby adjusting the relative position of the magnetic rod 42 and the magnetic rod sleeve 32.

[0261] When extracting nucleic acid, specific processes including mixing, magnetic adsorption and the like are performed. In the mixing stage, the magnetic rod sleeve 32 needs to move alone to mix the nucleic acid and magnetic beads; in the magnetic adsorption stage, the magnetic rod 42 is inserted into the magnetic rod sleeve 32 to keep the combination of the magnetic beads and the nucleic acid adsorbed on the surface of the magnetic rod sleeve 32, and the magnetic rod sleeve 32 and the magnetic rod 42 are synchronously displaced to transfer the combination of the magnetic beads and the nucleic acid to the subsequent washing and elution holes to complete the corresponding washing and elution processes. That is, in the above mixing stage, the magnetic rod sleeve 32 moves alone, and in the magnetic adsorption stage, the magnetic rod sleeve 32 and the magnetic rod 42 move synchronously.

[0262] Through the technical scheme provided in the present application, in the mixing stage, the lifting driving member 74 drives the magnetic rod sleeve 32 to displace. In the magnetic adsorption stage, the secondary driving member 43 first drives the magnetic rod 42 to be inserted into the magnetic rod sleeve 32, and keeps the magnetic rod 42 stationary relative to the magnetic rod sleeve 32 to adsorb the combination of the magnetic beads and the nucleic acid on the surface of the magnetic rod sleeve 32; and then the lifting driving member 74 drives the magnetic rod sleeve 32 and the magnetic rod 42 to synchronously move, thereby carrying the combination of the magnetic beads and the nucleic acid to transfer and perform subsequent washing and elution.

[0263] As can be seen, in the technical scheme of the present application, in the magnetic adsorption stage, the combination of the magnetic beads and the nucleic acid is adsorbed on the surface of the magnetic rod sleeve 32, and the lifting driving member 74 drives the magnetic rod sleeve 32 and the magnetic rod 42 to synchronously move, which, compared with the mode of driving the magnetic rod sleeve 32 and the magnetic rod 42 to move respectively by using separate driving structures, reduces the difficulty of operation and avoids the problems of equipment damage and unstable adsorption caused by the asynchronous movement of the magnetic rod sleeve 32 and the magnetic rod 42.

[0264] It should be noted that the driving member 21, the displacement driving member 72, the lifting driving member 74 and the secondary driving member 43 in the present application are only used for illustrative description in the above description, and in actual use, different driving structures can be selected and arranged as needed, as long as the corresponding driving functions can be stably achieved, and the purpose is to achieve the required functions.

[0265] Referring to FIG. 15, the nucleic acid extractor further comprises a controller 8, a photoelectric sensor 81 and a warning element 82, wherein the photoelectric sensor 81 is fixedly connected in the box body 1 and is selectively installed on the hatch 12 or the inner side wall of the box body 1, and is used to detect the relative position of the magnetic rod sleeve 32 relative to the magnetic rod sleeve frame 31; the controller 8 is in signal connection with the photoelectric sensor 81 and the warning element 82, and is used to trigger the warning element 82 to send an alarm signal when the photoelectric sensor 81 detects that the position of the magnetic rod sleeve 32 relative to the magnetic rod sleeve frame 31 is deviated.

[0266] The plurality of magnetic rod sleeves 32 are a group and are inserted on the magnetic rod sleeve frame 31. The position of the magnetic rod sleeve 32 is detected by the photoelectric sensor 81 to determine whether it is inserted in place. When it is detected that the magnetic rod sleeve 32 is not inserted in the specified position, the controller 8 triggers the warning element 82 to send an alarm signal, and the staff adjusts the position of the magnetic rod sleeve 32. In this way, the problem of collision between the magnetic rod sleeve 32 and the consumables 6 and collision between the magnetic rod 42 and the magnetic rod sleeve 32 caused by nucleic acid extraction in the case of not being inserted in the specified position can be avoided.

[0267] The warning element 82 can be a warning light, a buzzer, a display screen and the like, which prompts the staff when the alarm signal is triggered. When the alarm element is a display screen, the installation position of the magnetic rod sleeve 32 can also be displayed on the display screen in real time. If there is a position deviation during the initial installation of the magnetic rod sleeve 32, it can be corrected to the right position at one time, which is more convenient.

[0268] In addition, in actual setting, the controller 8 is also in signal connection with the driving member 21, the displacement driving member 72, the lifting driving member 74 and the secondary driving member 43. Of course, other sensors can also be set in signal connection with the controller 8. The controller 8 controls the actions of various components according to the set program or the information detected by the sensors. This part is the existing design in the related field, and the person skilled in the art can selectively set it according to the actual needs. This part will not be described in detail in this embodiment.

[0269] Referring to FIG. 16, the nucleic acid extractor further comprises a ventilation module 9, which forms an air duct of an experimental area for the extraction process in the box body 1. Specifically, the ventilation module 9 comprises an air inlet 91 and an air outlet 92. The air inlet 91 and the air outlet 92 are arranged to communicate the inside and outside of the box body 1. A fan 93 is arranged at the air outlet 92 to drive the flow of gas, forming an air flow from the air inlet 91 to the air outlet 92 and through the experimental area in the box body 1. The air inlet 91 and the air outlet 92 are located at two positions away from each other on the box body 1, and the height of the air inlet 91 is lower than that of the air outlet 92.

[0270] Exemplarily, the air inlet 91 is located at the lower side of one of the side walls of the box body 1 adjacent to the side on which the opening is located, the air outlet 92 is located at the upper side of the side wall of the box body 1 away from the opening, and the air outlet 92 is located on the side away from the side wall on which the air inlet 91 is located.

[0271] Through the above-mentioned air duct formed, the gas flow in the box body 1 is driven, so as to avoid vortex of the gas volatilized in the experiment process in the box body 1, thereby avoiding pollution of the gas sol to improve the purity of the extracted nucleic acid and the quality of the extraction.

[0272] In addition, a filter is arranged at the position of the air outlet 92 to filter the airflow in the box body 1, so as to avoid pollution to the outside.

[0273] In actual arrangement, the air outlet 92 can also be provided as two, and the fan 93 and the filter are arranged corresponding to the air outlet 92; for example, the two air outlets 92 are arranged on the side wall of the box body 1 away from the opening and arranged along the direction close to and away from the side wall on which the air inlet 91 is located. Alternatively, the air outlet 92 can also be arranged on the side wall away from the side wall on which the air inlet 91 is located.

[0274] The application also provides a library preparation device, which comprises the above-mentioned nucleic acid extractor, and the nucleic acid extractor is integrated into the library preparation device. Compared with the prior art in which the nucleic acid extractor is independently arranged, the use convenience is improved and the risk of pollution is reduced.

[0275] The application also provides a nucleic acid extraction method, which is realized based on the above-mentioned nucleic acid extractor and mainly comprises the following steps.

[0276] The door 12 is opened, and the placing table 2 is driven by the driving member 21 to slide from the reaction position in the box body 1 to the taking and placing position outside the box body 1;

[0277] The door 12 is closed;

[0278] The sample to be extracted is placed on the placing table 2, the door 12 is opened, and the placing table 2 loaded with the sample is driven by the driving member 21 to slide to the reaction position;

[0279] The door 12 is closed;

[0280] After the reaction is completed, the door 12 is opened, the placing table 2 loaded with the extracted nucleic acid is driven by the driving member 21 to slide to the taking and placing position, and the door 12 is closed.

[0281] Wherein, the sample to be extracted is placed on the placement table 2, which specifically includes the following steps: determining the number of samples that need to be extracted, installing the corresponding number of consumables 6 on the support frame 5 for the number of samples that need to be extracted, and placing samples and reagents required for extraction in each consumable hole 61 of the consumable 6. For the magnetic beads required in the process of extracting nucleic acid, the magnetic beads can be placed in one of the consumable holes 61, or a structure for placing magnetic beads can be additionally provided to store the magnetic beads. In the embodiments of the present application, the magnetic beads are placed in one of the consumable holes 61 as an example for illustration.

[0282] At this time, the placement table 2 is located inside the box body 1, and the opening of the box body 1 is in a closed state by the hatch 12. After opening the hatch 12, the driving member 21 drives the placement table 2 to move from the opening to the outside of the box body 1, and then the support frame 5 loaded with the consumables 6 is placed at the corresponding position on the placement table 2.

[0283] During the execution of the above steps, the placement table 2 is always located outside the box body 1, and the hatch 12 is in a closed state, keeping the inside of the box body 1 isolated from the outside.

[0284] After the above steps are completed, the hatch 12 is opened, the placement table 2 is slid to the reaction position inside the box body 1 by the driving member 21, and the hatch 12 is closed, so that the inside of the box body 1 is isolated from the outside, avoiding the problem of contamination caused by the diffusion of aerosol generated in the process of extracting nucleic acid to the external area.

[0285] After the placement table 2 loaded with the support frame 5 moves to the reaction position, the displacement driving member 72 drives the displacement plate 71 to slide in the second direction, so that the magnetic rod sleeve 32 moves to the position corresponding to the consumable hole 61 where the magnetic beads are placed; then the lifting driving member 74 drives the magnetic rod sleeve frame 31 to slide in the third direction, driving the magnetic rod sleeve 32 to move synchronously with the magnetic rod 42. During this process, the magnetic rod 42 remains stationary relative to the magnetic rod sleeve 32, and the two are in a state of mutual separation. The magnetic rod sleeve 32 is driven to extend into the consumable hole 61 where the magnetic beads are stored, and the magnetic rod sleeve 32 is driven to slide up and down reciprocally, mixing the magnetic beads. Then the secondary driving member 43 drives the magnetic rod 42 to slide relative to the magnetic rod sleeve 32, driving the magnetic rod 42 to insert into the corresponding magnetic rod sleeve 32, adsorbing the magnetic beads on the magnetic rod sleeve 32, and transferring the magnetic beads by the magnetic rod sleeve 32.

[0286] The lifting driving member 74 drives the magnetic rod sleeve 32 and the magnetic rod 42 to move out of the consumable hole 61 where the magnetic beads are stored. In the subsequent stage, the magnetic rod 42 and the magnetic rod sleeve 32 remain relatively stationary, stably adsorbing the magnetic beads on the surface of the magnetic rod sleeve 32. The magnetic rod 42 and the magnetic rod sleeve 32 are driven by the lifting driving member 74 and the displacement driving member 72 to move synchronously in the third direction and the second direction.

[0287] After the magnetic beads are adsorbed on the surface of the magnetic rod sleeve 32, the magnetic beads are moved to the sample position, the magnetic rod sleeve 32 carries the magnetic beads into the sample, the magnetic rod 42 is extracted from the magnetic rod sleeve 32 and remains separated, the magnetic rod sleeve 32 moves to shake and mix, and the sample is lysed; then the magnetic rod 42 moves and is inserted into the magnetic rod sleeve 32 and remains stationary with the magnetic rod sleeve 32, keeping the combination of the magnetic beads and the nucleic acid adsorbed on the magnetic rod sleeve 32. During the up-and-down shaking and mixing process, the temperature required for lysis is provided by the heating structure, which can be adjusted according to the actual operation of different samples. For example, the lysis temperature is adjusted to room temperature-120°C.

[0288] During actual shaking and mixing, the height of the magnetic rod sleeve 32 during mixing is set according to the volume of the different samples, and different mixing amplitudes are set to achieve the best mixing effect. How to calculate the mixing height and the shaking frequency during mixing according to the sample volume is determined by those skilled in the art based on known technology, and the determination method is not specifically introduced in this embodiment.

[0289] Then, the magnetic beads combined with the nucleic acid are adsorbed on the magnetic rod sleeve 32 and are transferred to the rinsing hole for up-and-down shaking and rinsing, and then the magnetic beads are moved to the air for drying. During this process, the magnetic beads can be rinsed multiple times according to actual needs to achieve better rinsing effect.

[0290] Finally, the rinsed magnetic beads are carried to the elution hole, so that the nucleic acid on the magnetic beads is eluted and falls into the elution hole, waiting for subsequent processes.

[0291] It should be noted that the above-mentioned lysis hole, rinsing hole and elution hole refer to the consumable hole 61 corresponding to lysis, rinsing and elution. The corresponding accommodation structure is provided at the corresponding hole position, and it is not simply an open hole structure. In order to facilitate description, it is simply referred to as a lysis hole, a rinsing hole and an elution hole.

[0292] In the above steps, the nucleic acid eluted is temporarily stored in the consumable hole 61 of the consumable 6, the hatch 12 is opened, the driving member 21 drives the placement table 2 to slide to the taking and placing position outside the box body 1, and the nucleic acid extracted is used in subsequent processes. After the placement table 2 moves to the taking and placing position, the hatch 12 is closed to seal the internal cavity of the box body 1, reducing the pollution problem with the surrounding area.

[0293] The quality control container is a test tube (such as a centrifugal tube) for quality control, and the accommodation position is a tube position seat for accommodating the test tube. The application also provides a test tube clamping device, which has the advantages of simple structure, small size and convenient opening, liquid adding and closing of the test tube by automatic equipment (i.e. quality control operation hand assembly).

[0294] The tube clamping device provided by the embodiments of the present application can be applied to a quality inspection device (i.e., a quality control system). When the nucleic acid extract carried in the tube is subjected to quality inspection, the tube clamping device can be used to clamp the tube, a mechanical hand (i.e., a quality control operation hand assembly) can be used to unscrew the tube cover, a reagent that can produce fluorescence when combined with the nucleic acid can be added, and the mechanical hand can be used to screw the tube cover again. The nucleic acid concentration can be detected by a fluorescence quantitative instrument through the fluorescence produced by the combination of the added reagent and the nucleic acid.

[0295] Referring to FIG. 17, which is a schematic diagram of the cooperation structure of the tube seat and the tube holder in the embodiments of the present application. In the embodiments, the tube clamping device includes a tube seat 310 and a tube holder 110 used for elastically clamping the tube 410. The tube holder 110 is used to directly elastically clamp the tube 410. The elastic clamping refers to that the part of the tube holder 110 clamping the tube 410 has a certain elasticity. When the tube 410 is inserted into the tube holder 110, the tube holder 110 can be expanded and produce a springback force, and the springback force is used to clamp the outer wall of the tube 410, so that the tube 410 is fixed in the tube holder 110. The elastic clamping can be achieved in various ways. For example, the part of the tube holder 110 clamping the tube 410 is made of elastic material. When the tube 410 is inserted into the tube holder 110, the elastic material is expanded by the support force provided by the outer wall of the tube 410. At this time, the tube holder 110 is elastically deformed and produces a contraction force towards the axial direction of the tube 410, so as to elastically clamp the tube 410. The elastic member can also be arranged along the circumferential direction of the tube holder 110. The elastic member directly or indirectly abuts against the outer wall of the tube 410, so as to elastically clamp the tube 410.

[0296] In the present application, the tube seat 310 is used to install the tube holder 110. The tube seat 310 is provided with a first accommodating cavity 314 with one end open. The tube holder 110 is slidingly assembled in the first accommodating cavity 314 and can slide along the axial direction of the first accommodating cavity 314. In order to realize the sliding of the tube holder 110 in the first accommodating cavity 314, the diameter of the first accommodating cavity 314 is slightly larger than the outer diameter of the tube holder 110. The cavity length of the first accommodating cavity 314 is greater than the length of the tube holder 110, so that the tube holder 110 can slide along the axial direction of the first accommodating cavity 314 during the opening and closing of the tube cap 411. It should be understood that in the embodiments, the height position of the tube cap 411 of the tube 410 does not change during the opening and closing of the tube cap 411. Since the tube cap 411 is connected with the tube 410 through threads, the tube cap 411 will rotate during the opening and closing of the tube cap 411. The tube 410 is driven by the threads to slide along the axial direction of the first accommodating cavity 314. The sliding distance is the thread length of the tube 410.

[0297] In order to make the test tube 410 and the cap 411 effectively contact in the process of opening and closing. Specifically, the first cavity 314 is provided with a resilient element, one end of the resilient element is in abutment with the tube seat 310, and the other end is in abutment with the tube holder 110. The tube holder 110 and the tube seat 310 are elastically connected along the axis direction of the tube seat 310 through the resilient element. In the process of opening the test tube 410, the mechanical hand holds the cap 411 of the test tube 410 and rotates, at this time the test tube 410 is driven by the thread to slide away from the cap 411 along the first cavity 314, the tube holder 110 is forced to slide along the axis of the tube seat 310 to the bottom of the first cavity 314, and the resilient element is compressed. Under the action of the resilient element, after the opening is completed, the test tube 410 returns to the original height. In the process of closing the test tube 410, the mechanical hand holds the cap 411 and rotates to the opening position, in this process, the test tube 410 will be pressed first, and the resilient element returns to the compressed state. And because the resilient element supports the tube holder 110 to provide a support force towards the cap 411, the test tube 410 has a tendency to move towards the cap 411, so that the test tube 410 is in abutment with the cap 411. When the mechanical hand holds the cap 411 and rotates, the test tube 410 and the cap 411 are screwed under the action of the thread. At this time, the test tube 410 adaptively moves towards the cap 411 along the thread, and the resilient element is released.

[0298] In order to facilitate the mechanical hand to perform the opening and closing operation on the test tube 410, the embodiment is provided with an anti-rotation structure on the side wall of the first cavity 314 for preventing the test tube 410 from rotating. Because the cap 411 is rotated by the mechanical hand in the process of opening and closing, in order to achieve opening and closing, it is necessary to keep the test tube 410 in a non-rotating state. The anti-rotation structure can be a concave-convex structure provided on the side wall of the first cavity 314 and the outer shape of the test tube 410 is clamped, which is designed to prevent the test tube 410 from rotating in the process of opening and closing.

[0299] As can be seen from the above description, in the embodiment disclosed in the present application, the tube holder 110 is slidably arranged in the tube seat 310, and the bottom of the tube holder 110 is elastically connected with the tube seat 310 through the resilient element. The test tube 410 is elastically clamped by the tube holder 110, and the tube holder 110 can adaptively slide in the tube seat 310 in the process of opening and closing the test tube 410. It has the advantages of small volume and simple structure.

[0300] As an optional implementation, referring to Figure 18, which is a three-dimensional structural diagram of the tube holder 110 of this application, the tube holder 110 has a second receiving cavity 116 for accommodating test tubes 410, with one end of the second receiving cavity 116 open. When inserting the test tube 410, the test tube 410 is inserted into the second receiving cavity 116 through the open end of the second receiving cavity 116. The shape of the second receiving cavity 116 matches the shape of the test tube 410, and is generally circular. To fix the test tube 410 inserted into the second receiving cavity 116, in this embodiment, a plurality of elastic retaining arms 11a are provided at the open end of the second receiving cavity 116. The multiple elastic retaining arms 11a surround the outer wall of the test tube 410, thus fixing the test tube 410.

[0301] It should be understood that, in this embodiment, the diameter of the inscribed circle of the plurality of elastic clamping arms 11a in their natural state is smaller than the diameter of the outer wall of the test tube 410. When the test tube 410 is inserted into the second receiving cavity 116, the plurality of elastic clamping arms 11a are spread open by the outer wall of the test tube 410. At the same time as being spread open, the plurality of elastic clamping arms 11a will also generate a rebound force toward the outer wall of the test tube 410, and the outer wall of the test tube 410 is elastically clamped by the rebound force.

[0302] To provide a uniform clamping force on the outer wall of the test tube 410, in this embodiment, multiple elastic clamping arms 11a are arranged at intervals around the axis of the second receiving cavity 116. The elastic clamping arms 11a can be arranged by creating slots 11b penetrating the cavity wall of the second receiving cavity 116, such as a straight, herringbone, or cross-shaped slot, dividing the cavity wall of the second receiving cavity 116 into multiple elastic clamping arms 11a. In this case, two adjacent elastic clamping arms 11a are not connected in the direction of the opening end of the second receiving cavity 116. When the test tube 410 is inserted, the multiple elastic clamping arms 11a undergo elastic deformation at the end face of the slot 11b, simultaneously generating a rebound force to elastically clamp the test tube 410. This application achieves elastic clamping of the test tube 410 by setting elastic clamping arms 11a on the tube support 110, resulting in a small size and simple structure.

[0303] Referring again to Figure 17, the bottom of the second receiving cavity 116 is a conical surface 113. At this time, the bottom surface of the test tube 410 is also set as a conical surface that matches the conical surface 113 at the bottom of the second receiving cavity 116. The two conical surfaces work together to position the test tube 410 and prevent the test tube 410 from being tilted in the second receiving cavity 116, which facilitates subsequent operations such as opening the cap, adding liquid, and closing the cap on the test tube 410.

[0304] As an optional embodiment, referring to FIG. 17 and FIG. 19, when the anti-rotation structure is the guide groove 123 arranged on the sidewall of the first accommodating cavity 314, the length direction of the guide groove 123 is arranged along the axis of the first accommodating cavity 314. And the side ears 412 are arranged on both sides of the test tube 410. After the test tube 410 is inserted into the tube holder 110, the side ears 412 on both sides of the test tube 410 are embedded in the guide groove 123. During the opening and closing of the test tube 410, due to the constant height of the tube cap 411, as the tube cap 411 rotates, the test tube 410 is driven by the thread to slide in the guide groove 123 in the first accommodating cavity 314. The anti-rotation structure in this embodiment is simple and occupies small space.

[0305] In order to limit the tube holder 110 in the first accommodating cavity 314 of the tube seat 310, the first accommodating cavity 314 is specifically arranged as a first cavity 314a and a second cavity 314b which are connected. The diameter of the first cavity 314a is smaller than the diameter of the second cavity 314b, so that a step surface 122 is formed between the first cavity 314a and the second cavity 314b, and the tube holder 110 is limited in the first accommodating cavity 314 by the step surface 122.

[0306] When the tube holder 110 is limited in the first accommodating cavity 314 by the step surface 122, the opening end of the first accommodating cavity 314 is located in the first cavity 314a, so that the first accommodating cavity 314 as a whole is in the shape of an inverted "T". At this time, the tube holder 110 is provided with a step structure 114 corresponding to the step surface 122 in the first accommodating cavity 314. As an example, referring to FIG. 17, the lower part of the tube holder 110 has a larger diameter than the upper part, and the step structure 114 is formed corresponding to the step surface 122 between the first cavity 314a and the second cavity 314b. By the step structure 114 and the step surface 122 being stuck, the tube holder 110 is limited in the second cavity 314b. The gap between the lower part of the tube holder 110 and the cavity wall of the second cavity 314b can be 0.1-0.5mm, such as 0.1mm or 0.2mm, 0.3mm, 0.4mm, etc. The tube holder 110 can slide in the first accommodating cavity 314 while preventing the tube holder 110 from swinging radially.

[0307] When the first accommodating cavity 314 is specifically provided with the first cavity 314a and the second cavity 314b, in order to facilitate the installation of the tube holder 110 in the first accommodating cavity 314, the tube seat 310 is provided in a split structure in the embodiment. Specifically, the seat body 311 is detachably fixedly connected with the support block 312. The seat body 311 is installed above the support block 312 and is connected by bolts or other components. The first cavity 314a is arranged in the seat body 311, and the second cavity 314b is arranged in the support block 312. The first cavity 314a and the second cavity 314b are coaxially arranged, and at this time, the step surface 122 of the first accommodating cavity 314 is located at the matching surface of the seat body 311 and the support block 312. When installing the tube holder 110, the lower large-diameter part of the tube holder 110 can be first embedded and installed in the second cavity 314b of the support block 312, and then the seat body 311 is sleeved outside the tube holder 110 through the first cavity 314a, which has the advantage of convenient installation. In order to ensure that the first cavity 314a and the second cavity 314b can be coaxial after installation, a pin hole can be arranged corresponding to the seat body 311 and the support block 312, and a positioning pin is inserted into the pin hole for positioning.

[0308] In an alternative embodiment, the elastic element is a compression spring 610. One end of the compression spring 610 abuts against the bottom surface of the tube holder 110, and the other end abuts against the bottom surface of the first accommodating cavity 314, and the compression spring 610 is in a compressed state, supporting the tube holder 110. During the process of closing the cap of the test tube 410 by the mechanical hand, the test tube 410 and the cap 411 are kept in an elastically abutting state, so that the threads between the test tube 410 and the cap 411 can be smoothly screwed.

[0309] In order to prevent the stability of the tube holder 110 during the opening and closing of the test tube 410, a guide column 313 cooperating with one end of the compression spring 610 is arranged on the bottom surface of the first accommodating cavity 314, and a ring groove 115 cooperating with the other end of the compression spring 610 is arranged on the tube holder 110. Specifically, one end of the compression spring 610 is sleeved outside the guide column 313, and the other end is embedded in the ring groove 115. The two ends of the elastic element are respectively positioned in the radial direction by the guide column 313 and the ring groove 115, so that when the tube holder 110 moves along the axis of the first accommodating cavity 314, the compression spring 610 is prevented from moving in the first accommodating cavity 314 under stress, causing the position of the tube holder 110 supported by the compression spring 610 to deviate.

[0310] In order to facilitate the use of the above-mentioned test tube clamping device in the laboratory, the application further provides a movement mechanism for driving the tube seat 310 to move. Referring to Figures 19 and 20, the tube seat 310 is detachably connected to the movement mechanism, and the movement mechanism drives the tube seat 310 to move, thereby driving the test tube 410 in the tube seat 310 to move. Since batch operations on the test tube 410 are required in the laboratory, such as opening the cap, adding liquid, closing the cap, etc. In order to cooperate with the mechanical hand to achieve high efficiency operation, the application drives the tube seat 310 to move through the movement mechanism. The movement form of the tube seat 310 driven by the movement mechanism can be linear translation, curved translation or reciprocating motion, etc. As long as it can meet the work requirements of the laboratory, it will not be described in detail here.

[0311] Referring to Figures 19 and 20, when the tube seat 310 is driven to displace by the movement mechanism, in order to ensure that the test tube 410 in the tube seat 310 can run smoothly, the movement mechanism of the application includes a cross beam 511, and a slide rail 512 is arranged on the cross beam 511. The tube seat 310 is slidably assembled on the slide rail 512, and the tube seat 310 is driven to slide along the slide rail 512 by a driving member. When the tube seat 310 is displaced by the movement mechanism, the tube seat 310 moves along the slide rail 512, which can ensure the stability of the test tube 410 in the tube seat 310. In the present embodiment, the shape of the cross beam 511 is not specifically limited. The main function of the cross beam 511 is to support the slide rail 512, and the slide rail 512 defines the displacement trajectory of the tube seat 310 and improves the stability of the test tube 410 during displacement.

[0312] When the tube seat 310 is slidably connected to the slide rail 512, the tube seat 310 can be provided with a structure matching the slide rail 512 to achieve the sliding connection with the slide rail 512. Alternatively, a sliding block can be fixedly connected to the tube seat 310, and the sliding block is slidably connected to the slide rail 512, thereby achieving the sliding connection of the tube seat 310 to the slide rail 512.

[0313] As an example, referring to FIG. 19 and FIG. 20, the driving member includes a conveyor belt assembly and a driving motor 515 driving the conveyor belt assembly. Specifically, a support frame 517 can be arranged at the lower part of one end of the cross beam 511, the driving motor 515 is fixedly connected below or above the support frame 517, and a pulley 516 is connected to the output end of the driving motor 515. A idler pulley 514 is arranged at the other end of the cross beam 511 opposite to the driving motor 515, and the idler pulley 514 is rotatably mounted on the cross beam 511 through a mounting plate 513. The pulley 516 and the idler pulley 514 are connected through a conveyor belt 518. At this time, the tube seat 310 is fixedly connected with the conveyor belt 518 in the conveyor belt assembly, and the specific connection manner between the tube seat 310 and the conveyor belt 518 can adopt any connection means in the prior art, as long as the tube seat 310 can move synchronously with the conveyor belt 518. When the driving motor 515 rotates, the output end of the driving motor 515 synchronously drives the pulley 516 to rotate, and under the driving of the pulley 516, the conveyor belt 518 runs between the pulley 516 and the idler pulley 514, and the tube seat 310 is driven to displace by the conveyor belt 518.

[0314] In order to improve work efficiency, in the embodiment, slide rails 512 are arranged on both sides of the cross beam 511, and the tube seat 310 is slidably connected to the slide rails 512 on both sides of the cross beam 511. When the driving member operates, the tube seat 310 on both sides of the cross beam 511 can be synchronously driven to operate, thereby improving efficiency.

[0315] In order to facilitate the understanding of the technical scheme of the present application, in the embodiment, the position of the tube seat 310 on the cross beam 511 close to the driving motor 515 is named as the first position (located in the working area of the quality control pipetting assembly), and the position of the tube seat 310 on the cross beam 511 close to the idler pulley 514 is named as the second position (located in the working area of the quality control operation hand assembly). The working scene of one of the test tube clamping devices of the present application is described in detail as follows in combination with FIG. 20:

[0316] 1. When the tube seat 310 is located at the second position, the robot puts the test tube 410 into the tube seat 310. The robot holds the cap 411 of the test tube 410 and rotates to perform the opening cover action on the test tube 410;

[0317] 2. After the test tube 410 is opened, the driving motor 515 rotates to drive the tube seat 310 to move to the first position by the conveyor belt 518, and at this time, the test tube 410 is located at the first position with the tube seat 310;

[0318] 3. The single-channel pump and other devices (i.e., the quality control pipetting assembly) add reagents to the test tube 410 located at the first position and opened;

[0319] 4. After the reagent is added, the drive motor 515 rotates in the opposite direction to that in step 2, and the conveyor belt 518 drives the tube holder 310 back to the second position. The mechanical hand-held tube cap 411 performs the capping action on the test tube 410;

[0320] 5. The robotic arm removes the closed test tube 410 and places a new test tube 410 into the tube holder 310 in the second position, and the cycle continues.

[0321] The above-described working method is only one of the working methods of the test tube clamping device of this application. As can be seen from the above working scenario, by using the test tube clamping device of this application in conjunction with a robotic arm, the opening, closing, and liquid addition of test tubes 410 in batch samples are automated, thereby improving work efficiency. Moreover, the test tube clamping device of this application has the advantages of small size, simple structure, and convenient operation.

[0322] This application also provides a quality inspection device, which includes the test tube clamping device described above and a robotic arm for loosening or tightening the caps of the test tubes.

[0323] As can be seen from the above description, in the embodiments disclosed in this application, a tube support 110 is slidably disposed within the tube holder 310, and the bottom of the tube support 110 is elastically connected to the tube holder 310 via an elastic element. The test tube 410 is elastically held by the tube support 110, and the tube support 110 can adaptively slide within the tube holder 310 during the opening and closing of the test tube 410. This design has the advantages of small size and simple structure.

[0324] One or more embodiments of this specification are intended to cover all such substitutions, modifications, and variations that fall within the broad scope of the appended claims. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of one or more embodiments of this specification should be included within the scope of protection of this disclosure.

[0325] The above are merely specific embodiments of this application, but the scope of protection of this application is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in this application should be included within the scope of protection of this application. Therefore, the scope of protection of this application should be determined by the scope of the claims.

Claims

1. A nucleic acid sequencing pre-processing workstation, characterized in that, The nucleic acid extraction system is configured to controllably extract nucleic acid molecules from a biological sample. The library construction system is configured to controllably prepare the nucleic acid molecules into a library for sequencing. The quality control system is configured to controllably detect the concentration of the nucleic acid extraction product, and according to the detection result and the library construction loading parameter, add a uniformization reagent to the nucleic acid extraction product; and / or, controllably detect the concentration of the library construction product, and according to the detection result, add a uniformization reagent to the library construction product and dilute it to the concentration required for nucleic acid sequencing. The quality control system comprises a quantitative analyzer, a quality control operation hand assembly, a conveying mechanism, a quality control pipetting assembly, a construction reaction container, a quality control container and a detection reagent container.

2. The nucleic acid sequencing pre-treatment workstation of claim 1, wherein, The quality control container is configured to accommodate the nucleic acid extraction product or the library construction product to be detected, and can be placed in the quantitative analyzer to realize concentration detection. The detection reagent container is configured to accommodate one or more detection reagents required for concentration detection by the quantitative analyzer and reagents for adjusting the concentration of the nucleic acid extraction product and / or the library construction product. The conveying mechanism is configured to reciprocate the quality control container between the working area of the quality control operation hand assembly and the working area of the quality control pipetting assembly. The quality control operation hand assembly is configured to take the quality control container from the storage area of the quality control container, place it on the conveying mechanism, take the quality control container from the conveying mechanism and place it back in the storage area of the quality control container, and take the quality control container from the storage area of the quality control container and place it into the quantitative analyzer, and take the quality control container that has completed detection from the quantitative analyzer and place it back in the storage area of the quality control container. The quality control pipetting assembly is configured to suck a certain amount of detection reagent from the detection reagent container and add it to the quality control container, and suck a certain amount of reagent for adjusting the concentration of the nucleic acid extraction product and / or the library construction product and add it to the construction reaction container. The construction reaction container is configured to accommodate the nucleic acid extraction product after quality control and provide a reaction space for the library construction process. The library construction system comprises a PCR instrument, a library construction operation hand assembly, a library construction pipetting assembly, a library construction reagent container, a shaking and heating assembly and a magnet assembly.

3. The nucleic acid sequencing pre-treatment workstation of claim 2, wherein, The library construction operation hand assembly is configured to take the construction reaction container containing the nucleic acid extraction product after quality control from the quality control system, place the construction reaction container in the reagent loading area in the library construction system, and send the construction reaction container loaded with reagents to the PCR instrument. And / or, send the construction reaction container containing the library construction product to the quality control system for quality control. The library construction reagent container is configured to accommodate one or more library construction reagents required for library construction. The magnet assembly is configured to provide the necessary magnetic field environment for magnetic bead purification. The shaking and heating assembly is configured to provide a reaction space and a reaction temperature for magnetic bead purification during the library construction process. ​ The library construction pipetting assembly is configured to add library construction reagents from the library construction reagent container to the construction reaction container.

4. The nucleic acid sequencing pre-treatment workstation of claim 3, wherein, A waste container for accommodating the waste pipetting heads of the library construction pipetting assembly is further included.

5. The nucleic acid sequencing pre-treatment workstation of any one of claims 1 to 4, wherein, A rack is further included, and a workbench is arranged on the rack, and the nucleic acid extraction system, the quality control system, and the library construction system are arranged on the workbench.

6. The nucleic acid sequencing pre-treatment workstation of claim 5, wherein, When the library construction system includes a library construction manipulator assembly and a library construction pipetting assembly, the library construction manipetting assembly and the library construction pipetting assembly are arranged on the same three-axis motion platform, and the three-axis motion platform is arranged on the rack.

7. The nucleic acid sequencing pre-treatment workstation of claim 6, wherein, An information acquisition module arranged on the three-axis motion platform is further included, and the information acquisition module is used to acquire information of consumables in the nucleic acid sequencing pretreatment workstation.

8. The nucleic acid sequencing pre-treatment workstation of claim 5, wherein, An air filtering device is further included, and the air filtering device is arranged on the top of the rack.

9. The nucleic acid sequencing pre-treatment workstation of claim 2, wherein, The nucleic acid extraction system includes a box with a cavity formed inside, and the box is composed of a bottom plate, a side plate, and a top plate; The nucleic acid extraction system further includes a placement table and a magnetic rod sleeve assembly and a magnetic rod assembly arranged in the cavity; wherein the placement table is used to carry biological samples to be extracted and reagents required for extraction, and the magnetic rod sleeve assembly and the magnetic rod assembly are used to carry biological samples between different extraction processes; An opening is formed in the side plate for the placement table to pass through; A driving member arranged on the lower side of the bottom plate is further included, and a communication hole is formed in the bottom plate along a first direction, the driving member is connected with the placement table through the communication hole, and the placement table is moved between a taking and placing position and a reaction position along the first direction by the driving member; A hatch for opening and closing the opening is arranged on the side plate, and the opening is closed when the placement table is located at the taking and placing position or the reaction position; The taking and placing position is located outside the cavity, and the reaction position is located inside the cavity.

10. The nucleic acid sequencing pre-treatment workstation of claim 9, wherein, A sealing strip for sealing the communication hole is arranged on the bottom plate.

11. The nucleic acid sequencing pre-treatment workstation of claim 10, wherein, A rolling member is arranged on the side of the placement table facing the driving member, and the placement table is connected with the driving member through the rolling member; The sealing strip and the rolling member cooperate to jointly block the communication hole.

12. The nucleic acid sequencing pre-treatment workstation of claim 11, wherein, The rolling member includes a support block, a connecting block, and a plurality of limiting rollers; Two support blocks are arranged for connection with the driving member and the placement table; The connecting block and the limiting block are located between the two support blocks, and both ends of the connecting block and the limiting roller are connected to the support blocks; The sealing strip crosses the connecting block, and the limiting rollers are distributed on both sides of the connecting block to limit the sealing strip to keep cooperating with the rolling member to seal the communication hole.

13. The nucleic acid sequencing pre-treatment workstation of any one of claims 9-12, wherein, The driving member includes a driving lead screw rotatably connected with the bottom plate, a sliding block sleeved on the driving lead screw, a driving motor with an output end connected with the driving lead screw, and a frame fixedly connected on the lower side of the bottom plate; the driving lead screw and the driving motor are connected with the frame; The sliding block is threadedly connected with the driving lead screw and is fixedly connected with the placement table.

14. The nucleic acid sequencing pre-treatment workstation of claim 9, wherein, The support frame is carried on the storage table; The plurality of consumables are arranged on the support frame along a first direction and are detachably connected with the support frame; Each of the consumables is provided with a plurality of consumable holes along a second direction, and the plurality of consumable holes are respectively used for storing samples to be extracted and reagents required for extraction; The first direction is perpendicular to the second direction. The bottom plate is fixedly connected with a mounting frame, and the mounting frame is provided with a displacement assembly; 15. The nucleic acid sequencing pre-treatment workstation of claim 9, wherein, The displacement assembly includes a displacement plate slidably connected with the mounting frame along a second direction and a displacement driving element arranged on the mounting frame, and the displacement driving element is connected with the displacement plate to drive the displacement plate to slide; The magnetic rod sleeve assembly and the magnetic rod assembly are arranged on the displacement plate; The first direction is perpendicular to the second direction. The magnetic rod sleeve assembly includes a magnetic rod sleeve frame and a plurality of magnetic rod sleeves arranged on the magnetic rod sleeve frame, and the magnetic rod sleeve frame is slidably connected with the displacement plate along a third direction; 16. The nucleic acid sequencing pre-treatment workstation of claim 15, wherein, The displacement plate is further provided with a lifting driving element for driving the magnetic rod sleeve frame to slide relative to the displacement plate; The magnetic rod assembly includes a magnetic rod frame and a plurality of magnetic rods arranged on the magnetic rod frame, and the magnetic rod frame is slidably connected with the magnetic rod sleeve frame along the third direction; The magnetic rod frame is provided with a secondary driving element for driving the magnetic rod frame to slide relative to the magnetic rod sleeve frame; The third direction is perpendicular to the bottom plate, and the plurality of magnetic rods correspond to the plurality of magnetic rod sleeves one by one. The secondary driving element includes a screw rod threadedly matched with the magnetic rod sleeve frame and a motor fixed relative to the magnetic rod frame, and the motor is used to drive the screw rod to rotate; 17. The nucleic acid sequencing pre-treatment workstation of claim 16, wherein, The screw rod penetrates through the magnetic rod frame and is rotationally connected with the magnetic rod frame, and the magnetic rod frame and the screw rod are relatively fixed in the third direction. Further comprising a controller, a photoelectric sensor and a warning element; 18. The nucleic acid sequencing pre-treatment workstation of claim 17, wherein, The photoelectric sensor is arranged in the box and is used for detecting the position of the magnetic rod sleeve relative to the magnetic rod sleeve frame; The controller is signal connected with the photoelectric sensor and the warning element, and is used to trigger the warning element to send an alarm signal when the photoelectric sensor detects that the position of the magnetic rod sleeve relative to the magnetic rod sleeve frame is deviated. The support frame is detachably connected with a pressing element; 19. The nucleic acid sequencing pre-treatment workstation of claim 14, wherein, The pressing element and the support frame cooperate to clamp the consumable to limit the position of the consumable. The consumable is provided with an identification mark for identifying a biological sample on the consumable.

20. The nucleic acid sequencing pre-treatment workstation of claim 14, wherein, The identification mark is an identification code for scanning by an identification device, and the identification device is the information acquisition module.

21. The nucleic acid sequencing pre-treatment workstation of claim 20, wherein, The box is provided with a plurality of ultraviolet lamps.

22. The nucleic acid sequencing pre-treatment workstation of claim 9, wherein, The nucleic acid extraction system further comprises a ventilation module, and the ventilation module comprises an air inlet, an air outlet and a fan arranged on the box; 23. The nucleic acid sequencing pre-treatment workstation of claim 9, wherein, The air inlet and the air outlet are located at two positions away from each other on the box; The internal space of the box is communicated with the outside through the air inlet and the air outlet; The fan is located at the air outlet, forming an air flow from the air inlet to the air outlet and flowing through the experimental area in the box. ​ 24. The nucleic acid sequencing pre-treatment workstation of claim 23, wherein, The ventilation module further comprises a filter arranged at the air outlet for filtering the airflow.

25. The nucleic acid sequencing pre-treatment workstation of claim 2, wherein, The conveying mechanism comprises a conveying belt and at least two accommodation positions for accommodating the quality control containers fixed on the conveying belt; wherein, The conveying belt is configured to reciprocally convey the quality control containers between the working area of the quality control operation hand assembly and the working area of the quality control pipetting assembly.

26. The nucleic acid sequencing pre-treatment workstation of claim 25, wherein, The quality control containers are test tubes for quality control, and the accommodation positions are tube seat positions for accommodating the test tubes; The conveying mechanism further comprises a tube holder for elastically clamping the test tube; wherein, The tube seat position is provided with a first accommodation cavity with an open end; the tube holder is slidingly assembled in the first accommodation cavity and can slide along the axis direction of the first accommodation cavity; The first accommodation cavity is provided with an elastic element, one end of which is in abutment with the tube seat position, and the other end is in abutment with the tube holder; The side wall of the first accommodation cavity is provided with an anti-rotation structure for preventing the test tube from rotating.

27. The nucleic acid sequencing pre-treatment workstation of claim 26, wherein, The tube holder has a second accommodation cavity for accommodating the test tube, the second accommodation cavity has an open end, and the open end of the second accommodation cavity is provided with a plurality of elastic clamping arms, which are arranged around the axis of the second accommodation cavity.

28. The nucleic acid sequencing pre-treatment workstation of claim 27, wherein, The bottom of the second accommodation cavity is a tapered surface.

29. The nucleic acid sequencing pre-treatment workstation of claim 26, wherein, The anti-rotation structure is a guide groove provided on the side wall of the first accommodation cavity.

30. The nucleic acid sequencing pre-treatment workstation of claim 29, wherein, The first accommodation cavity comprises a first cavity and a second cavity in communication, and the open end of the first accommodation cavity is located in the first cavity; wherein, a stepped surface is formed between the first cavity and the second cavity; The tube holder has a stepped structure that limits the cooperation with the stepped surface of the first accommodation cavity.

31. The nucleic acid sequencing pre-treatment workstation of claim 30, wherein, The tube seat position comprises a seat body and a support block detachably fixedly connected with the seat body; The first cavity is located in the seat body, and the second cavity is located in the support seat.

32. The nucleic acid sequencing pre-treatment workstation of claim 26, wherein, The elastic element is a compression spring; The bottom surface of the first accommodation cavity is provided with a guide column matched with one end of the compression spring; The tube holder is provided with a ring groove matched with the other end of the compression spring.

33. The nucleic acid pre-processing workstation of any one of claims 25-32, wherein, Further comprising a movement mechanism for driving the movement of the tube seat position.

34. The nucleic acid pre-processing workstation of claim 33, wherein, The movement mechanism comprises a cross beam and a slide rail provided on the cross beam; the tube seat position is slidingly assembled in the slide rail; the movement mechanism further comprises a driving member for driving the sliding of the tube seat position.

35. The nucleic acid pre-processing workstation of claim 34, wherein, The quality control operation hand assembly is further configured to loosen the cap of the test tube after taking out the quality control container from the storage area of the quality control container and placing it on the conveying mechanism, or tighten the cap of the test tube before taking out the quality control container from the conveying mechanism and placing it back to the storage area of the quality control container.

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