CN114134032A

CN114134032A - Gene sequencing pretreatment device

Info

Publication number: CN114134032A
Application number: CN202111458753.3A
Authority: CN
Inventors: 俞郴荣; 徐涛; 骆志成; 樊伟东; 骆广进
Original assignee: HANGZHOU ALLSHENG INSTRUMENTS CO Ltd
Current assignee: HANGZHOU ALLSHENG INSTRUMENTS CO Ltd
Priority date: 2021-12-02
Filing date: 2021-12-02
Publication date: 2022-03-04
Anticipated expiration: 2041-12-02
Also published as: CN114134032B

Abstract

The application relates to a processing apparatus before gene sequencing, the processing apparatus before gene sequencing of this application includes: the system comprises a rack, a kit, a pipettor, a main control module, a station switching module and a temperature control module, wherein the rack is provided with at least one magnetic suction module; the kit is arranged in the rack, and a plurality of functional hole sites are arranged on the kit; the pipettor is arranged in the rack; the station switching module is arranged on the rack and used for driving the pipettor and/or the kit so as to transfer the pipettor among the functional hole sites; the temperature control module is arranged on the rack, the kit and/or the pipettor and used for adjusting the temperature; the magnetic suction module is arranged on the kit and is positioned at one functional hole position and used for controlling magnetic suction; the main control module is connected with the station switching module, the temperature control module and the magnetic suction module. Therefore, the method and the device can realize the integrated functions of nucleic acid extraction, library construction and PCR amplification, have high automation degree, reduce the operation difficulty and improve the working efficiency.

Description

Gene sequencing pretreatment device

Technical Field

The application relates to the field of gene detection, in particular to a gene sequencing pretreatment device.

Background

Gene detection has become a main research means used in clinical molecular experiments, for example, the diagnosis and suspected resolution of patients infected by the novel coronavirus, infection of postoperative severe patients and the like need to be assisted by the gene detection result. The precondition of gene detection is the extraction of biological sample nucleic acids of high quality and purity. Therefore, nucleic acid extraction and purification, gene library construction and PCR amplification become important steps in gene detection.

In the prior art, most of gene pretreatment processes such as nucleic acid extraction, gene library construction and the like are manually operated under an open experimental environment or all the steps are finished by means of instruments, so that the operation is troublesome, the working efficiency is low and the automation degree is low.

Disclosure of Invention

The utility model aims at providing a processing apparatus before gene sequencing, its degree of automation is high.

The embodiment of the application is realized as follows:

in one aspect, the present application provides a gene sequencing pretreatment apparatus, including: the device comprises a rack, a kit, a pipettor, a main control module, a station switching module, a temperature control module and at least one magnetic suction module; the kit is arranged in the rack, and a plurality of functional hole sites are arranged on the kit; the pipettor is arranged in the rack; the station switching module is arranged on the rack and used for driving the pipettor and/or the kit so as to transfer the pipettor among the functional hole sites; the temperature control module is arranged on the rack, the kit and/or the pipettor and used for adjusting the temperature; the magnetic suction module is arranged on the kit and is positioned at one functional hole position and used for controlling magnetic suction; the main control module is connected with the station switching module, the temperature control module and the magnetic suction module.

In one embodiment, each magnetic module comprises: the magnetic attraction driving mechanism is connected with the magnetic part and used for controlling the working state of the magnetic part.

In one embodiment, two magnetic members are provided, namely a cylindrical magnetic member and an annular magnetic member, and the cylindrical magnetic member is used for bottom surface magnetic attraction on the bottom of the functional hole; the annular magnetic part is sleeved outside the cylindrical magnetic part and used for performing annular magnetic attraction on the bottom of the functional hole site; the magnetic attraction driving mechanism is connected with the cylindrical magnetic part and the annular magnetic part and is used for controlling the working states of the cylindrical magnetic part and the annular magnetic part.

In one embodiment, the magnetic member is a permanent magnet, and the magnetic driving mechanism includes: the magnetic attraction type belt wheel assembly comprises a belt wheel assembly, a magnetic attraction driving motor, a first lifting plate, a second lifting plate and at least one sliding rail, wherein the belt wheel assembly comprises a transmission belt, and a first shifting block and a second shifting block which are opposite to each other are arranged on the transmission belt; the magnetic attraction driving motor is in transmission connection with the belt wheel assembly and is used for driving the transmission belt to rotate so as to control the first shifting block and the second shifting block to lift; the first lifting plate is movably arranged on the sliding rail and is connected with the annular magnetic part and the first shifting block; the second lifting plate can be movably arranged on the sliding rail and is connected with the cylindrical magnetic part and the second shifting block.

In one embodiment, the magnetic driving mechanism includes: spacing subassembly, spacing subassembly are including the first piece and the second piece of attracting of mutually supporting, and the second lifter plate is located to first piece of attracting, and the slide rail is located to the second piece of attracting.

In one embodiment, the magnetic driving mechanism includes: the resetting component is arranged on the sliding rail and used for resetting the first lifting plate and the second lifting plate. Wherein the return assembly may be a spring.

In one embodiment, the belt wheel assembly further comprises a transmission wheel, and the magnetic attraction driving motor is in transmission connection with the belt wheel assembly through the transmission wheel.

In one embodiment, there are two slide rails, and the first lifting plate and the second lifting plate are respectively disposed on two different slide rails.

In an embodiment, there are two slide rails, and both the two slide rails are connected to the first lifting plate and the second lifting plate, and the second lifting plate is located below the first lifting plate.

In one embodiment, the number of the slide rails is 1, and the first lifting plate and the second lifting plate are both arranged on the same slide rail.

In one embodiment, the magnetic member is an electromagnet, and the magnetic driving mechanism is a control circuit.

In one embodiment, the pretreatment apparatus for gene sequencing further comprises: and the environment control module is arranged in the rack, is connected with the main control module and is used for adjusting the environment condition in the rack.

In one embodiment, the environmental control module includes: and the at least one ultraviolet light source is arranged on the frame.

In one embodiment, the environmental control module includes: at least one filter piece, be equipped with a plurality of vents in the frame, filter the piece and locate the vent.

In one embodiment, the environmental control module includes: and the ventilation mechanism is arranged in the rack, is matched with the plurality of ventilation openings and is used for controlling the wind pressure in the rack.

In one embodiment, the ventilation mechanism comprises at least one wind pressure monitor, at least one wind inlet member and at least one wind outlet member.

In one embodiment, the station switching module includes: the device comprises a first platform, an XZ direction movement mechanism, a second platform and a Y direction movement mechanism, wherein the first platform is connected with a liquid transfer device; the XZ direction movement mechanism is connected with the first platform and is used for enabling the first platform to move along the X direction or the Z direction; the second platform is connected with the kit; the Y-direction movement mechanism is connected with the second platform and used for enabling the second platform to move along the Y direction.

In one embodiment, the X-direction, the Z-direction and the Y-direction are perpendicular to each other.

In one embodiment, the XZ-direction movement mechanism includes: the X-direction lead screw is arranged on the rack along the X direction; the X-direction driving motor is in transmission connection with the X-direction lead screw and is connected with a connecting piece, and the X-direction driving motor is used for driving the X-direction lead screw to rotate so as to enable the X-direction driving motor and the connecting piece to move along the X direction; the Z-direction lead screw is arranged on the connecting piece along the Z direction, is in transmission connection with the first platform and is used for driving the first platform to move along the Z direction; the Z-direction driving motor is in transmission connection with the Z-direction lead screw and can drive the Z-direction lead screw to rotate.

In one embodiment, the XZ-direction moving mechanism further includes: the Z-direction guide rail is arranged on the connecting piece along the Z direction and is provided with at least one Z-direction sliding block; the Z-direction lead screw is connected with the connecting piece through the Z-direction guide rail, and a nut of the Z-direction lead screw is connected with the Z-direction sliding block.

In one embodiment, the XZ-direction moving mechanism further includes: and a synchronous belt transmission component. The synchronous belt transmission assembly comprises two synchronous belt wheels and a synchronous belt, one synchronous belt wheel is connected with the Z-direction driving motor, and the other synchronous belt wheel is connected with the Z-direction lead screw. Wherein, Z direction motor and Z direction lead screw locate the both sides of Z direction guide rail respectively, and the top of Z direction motor and Z direction lead screw is located to the synchronous belt drive assembly.

In one embodiment, the Y-direction movement mechanism includes: the Y-direction lead screw is arranged on the rack along the Y direction; the Y-direction driving motor is in transmission connection with the Y-direction lead screw and can drive the Y-direction lead screw to rotate; the Y-direction lead screw is in transmission connection with the second platform and used for driving the second platform to move along the Y direction, the Y-direction guide rail is arranged on the rack along the Y direction, at least one Y-direction sliding block is arranged on the Y-direction guide rail, and the Y-direction sliding block is connected with the second platform.

In one embodiment, a pipette comprises: the liquid transferring cavity is connected with a liquid transferring gun head; the liquid-transferring driving piece is connected with the liquid-transferring cavity and used for controlling liquid transfer of the liquid-transferring gun head.

In one embodiment, the pipette further comprises: move liquid detection piece, locate and move liquid the cavity for detect move liquid atmospheric pressure and/or temperature of cavity.

In one embodiment, the pipette further comprises: and the suction head withdrawing mechanism comprises a suction head withdrawing movement plate which is configured to move along the axis of the pipette head.

In one embodiment, the temperature control module includes: the hot lid heating unit is arranged in the rack and positioned above the kit and used for heating the hot lid.

In one embodiment, the thermal cover heating unit comprises a heating column and a thermal cover annealing mechanism, wherein the heating column is used for heating the thermal cover; the thermal cover moving plate is configured to be movable along an axis of the heating column.

In one embodiment, the heating column, the suction head withdrawing mechanism and the heat lid withdrawing mechanism are all arranged on the first platform.

In one embodiment, the moving plate of the de-suction head is connected with the moving plate of the de-heat cover, and the moving plate of the de-suction head and the moving plate of the de-heat cover are both in transmission connection with the lifting driving assembly.

In one embodiment, the plurality of functional holes includes: at least one tip access well, at least one thermal cover access well, at least one tube access well, at least one first reaction well, at least one second reaction well, and a plurality of reagent wells.

In one embodiment, the temperature control module includes: the reagent zone temperature control unit is arranged at the bottom of the reagent kit and is used for controlling the temperature of materials in the reagent holes; the reaction area temperature control unit is arranged at the bottom of the kit and is used for controlling the temperature of the materials in the first reaction hole and the second reaction hole.

In one embodiment, the reagent zone temperature control unit comprises: the heat-insulation device comprises a first heat-transfer block, a first heat-insulation sleeve, a first semiconductor refrigeration sheet, a first radiator and a first water absorption piece; the first heat transfer block is arranged at the bottom of the kit; the first heat-insulating sleeve is sleeved outside the first heat-transfer block; the first semiconductor refrigeration piece is provided with a first cold surface and a first hot surface, and the first cold surface is connected with the first heat transfer block; the first radiator is arranged on the first hot surface; the first water absorbing piece is arranged between the first heat transfer block and the first radiator and outside the first semiconductor refrigeration piece.

In one embodiment, the temperature control unit of the reaction zone comprises: and the heating sleeve is arranged at the bottom of the reagent kit and is provided with a first hole for accommodating the second reaction hole.

In one embodiment, the temperature control unit further comprises: the second heat transfer block is arranged at the bottom of the kit and is connected with the heating sleeve, and a second hole for accommodating the first reaction hole is formed in the second heat transfer block; the second heat-insulating sleeve is sleeved outside the second heat-transfer block; the second semiconductor refrigerating piece is provided with a second cold surface and a second hot surface, and the second cold surface is connected with the second heat transfer block; the second radiator is arranged on the second hot surface; the second water absorbing piece is arranged between the second radiator and the second heat transfer block and outside the second semiconductor refrigerating sheet.

In one embodiment, the second stage comprises: the kit comprises a first substrate and a second substrate, wherein the first substrate is connected with the kit; the second substrate is connected with the first substrate through a plurality of connecting rods and is arranged in parallel with the first substrate; the second substrate is provided with a shunting hole, the first radiator and the second radiator are arranged in the shunting hole, air inlets of the first radiator and the second radiator are located between the first substrate and the second substrate, and air outlets of the first radiator and the second radiator are located on one side, deviating from the first substrate, of the second substrate.

In one embodiment, the temperature control module includes: a plurality of temperature sensor, a plurality of temperature sensor locate frame, kit and/or pipettor.

In one embodiment, the reagent kit is hinged with a flip cover containing holes, so that the reagent kit is in an open structure.

Compared with the prior art, the beneficial effects of the application are that:

this application can control station switch module so that the pipettor shifts between each function hole site through host system, can also control magnetism to inhale the module and adjust magnetism, can also control temperature control module and carry out temperature regulation to can realize the integration function of set nucleic acid extraction, library construction and PCR amplification, degree of automation is higher, has reduced the operation degree of difficulty, and has improved work efficiency.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are required to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present application and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained from the drawings without inventive effort.

FIG. 1 is a schematic view of the structure of a pretreatment apparatus for gene sequencing according to an embodiment of the present invention.

FIG. 2 is an exploded view of a part of the structure of a pretreatment device for gene sequencing according to an embodiment of the present invention.

FIG. 3 is an exploded view of a part of the structure of a pretreatment device for gene sequencing according to an embodiment of the present invention.

FIG. 4 is a sectional view of a pretreatment apparatus for gene sequencing according to an embodiment of the present application.

FIG. 5 is a schematic view of a partial configuration of a pretreatment device for gene sequencing according to an embodiment of the present invention.

FIG. 6 is a schematic view of the connection of a pipette tip to a tip according to an embodiment of the present application.

FIG. 7 is a schematic diagram of a temperature control unit for a reagent zone according to an embodiment of the present application.

FIG. 8 is a cross-sectional view of a reagent zone temperature control unit according to one embodiment of the present application.

FIG. 9 is a schematic structural diagram of a temperature control unit of a reaction zone according to an embodiment of the present application.

FIG. 10 is a cross-sectional view of a temperature control unit for a reaction zone according to an embodiment of the present application.

FIG. 11 is a schematic view of a partial configuration of a pretreatment device for gene sequencing according to an embodiment of the present invention.

FIG. 12 is a schematic view of a partial configuration of a pretreatment device for gene sequencing according to an embodiment of the present invention.

FIG. 13 is a partial cross-sectional view of a pre-treatment apparatus for gene sequencing according to an embodiment of the present invention, in which a magnetic attraction module is in a loop-attraction state.

FIG. 14 is a partial cross-sectional view of a pre-treatment apparatus for gene sequencing according to an embodiment of the present invention, in a state where a magnetic attraction module is in a bottom attraction state.

Fig. 15 is a schematic structural view of a magnetic module in an initial state according to an embodiment of the present application.

Fig. 16 is a schematic structural view of a magnetic module in a ring-attraction state according to an embodiment of the present application.

Fig. 17 is a schematic structural view of a magnetic module in a bottom-attraction state according to an embodiment of the present application.

Fig. 18 is a top view of a kit according to an embodiment of the present application.

Icon: 1-a gene sequencing pretreatment device; 100-a frame; 110-a vent; 200-kit; 210-functional hole sites; 211-thermal lid storage hole; 212-a test tube storage well; 213-first reaction well; 214-second reaction well; 215-reagent well; 216-a tip access aperture; 217-a flip cover with holes; 300-a pipette; 310-pipetting chamber; 311-pipette tips; 320-pipetting driver; 330-a pipetting probe; 340-a suction head withdrawing mechanism; 341-moving plate of suction head; 400-station switching module; 410-a first platform; 420-XZ direction movement mechanism; a 421-X direction screw; 4211-X direction guide rail slide block assembly; a 422-X direction driving motor; 423-connecting piece; 424-Z direction guide rails; 425-Z direction slider; 426-Z direction lead screw; 427-Z direction driving motor; 429-synchronous belt drive assemblies; 430-a second platform; 431-a first substrate; 432-a second substrate; 433-a shunt hole; 434-air inlet; 435-air outlet; 436-connecting rods; 440-Y direction movement mechanism; 441-Y direction guide rails; a 442-Y direction slider; 443-Y direction screw; a 444-Y direction drive motor; 500-temperature control module; 510-a thermal lid heating unit; 511-a heating column; 512-a cover-annealing mechanism; 5121-moving plate of annealing cover; 514-a lift drive assembly; 520-reagent zone temperature control unit; 521-a first heat transfer block; 522-first thermal insulation sleeve; 523-first semiconductor refrigerating sheet; 524-a first heat sink; 526-the first absorbent member; 530-temperature control unit of reaction zone; 531-second heat transfer block; 5311-a second well; 532-second insulating sleeve; 533-second semiconductor chilling plate; 534-a second heat sink; 536-a second absorbent member; 537-heating jacket; 5371-a first aperture; 501-temperature sensor; 600-a magnetic module; 610-a magnetic member; 611-a cylindrical magnetic member; 612-ring-shaped magnetic member; 630-magnetic attraction drive mechanism; 631-a pulley assembly; 6311-driving belt; 6312-a drive wheel; 6313-first paddle; 6314-second paddle; 632-a magnetic attraction drive motor; 633-a first lifter plate; 634-a second lifter plate; 635-slide rail; 636-a limit component; 6361-first suction element; 6362-second attraction; 637-first fixing plate; 638-second fixing plate; 639-guide bar; 700-an environment control module; 710-a source of ultraviolet light; 720-a filter element; 730-a ventilation mechanism; 731-air intake member; 732-an air outlet member; 800-a main control module; 910-a thermal cover; 920-suction head.

Detailed Description

The terms "first," "second," "third," and the like are used for descriptive purposes only and not for purposes of indicating or implying relative importance, and do not denote any order or order. Furthermore, the terms "horizontal", "vertical", "overhang" and the like do not imply that the components are required to be absolutely horizontal or overhang, but may be slightly inclined. For example, "horizontal" merely means that the direction is more horizontal than "vertical" and does not mean that the structure must be perfectly horizontal, but may be slightly inclined. In the description of the present application, it should be noted that the terms "inside", "outside", "left", "right", "upper", "lower", and the like indicate orientations or positional relationships based on orientations or positional relationships shown in the drawings or orientations or positional relationships that are conventionally arranged when products of the application are used, and are used only for convenience in describing the application and simplifying the description, but do not indicate or imply that the referred device or element must have a specific orientation, be constructed in a specific orientation, and be operated, and thus, should not be construed as limiting the application. In the description of the present application, unless expressly stated or limited otherwise, the terms "disposed," "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements.

The gene detection is a medical detection technology, which extracts nucleic acid in peripheral venous blood, tissues and other body fluids of a detected person, analyzes DNA molecule or RNA molecule information in cells of the detected person through a detection device, so as to know the gene information of the detected person and further determine the cause of disease or the risk of disease. It should be understood that the gene detection is not limited to the detection of human body, but also includes the gene detection of animal and plant.

The nucleic acid extraction is a pretreatment process of gene detection, specific primers and probe design are carried out according to the sequence of known nucleic acid, the designed primers are synthesized, the extracted nucleic acid is used as a template to carry out a fluorescence quantitative PCR experiment, and the negative and positive of a target sample are judged according to a fluorescence signal. Nucleic acid extraction is a key step of gene detection, and the quality of the obtained nucleic acid directly influences the success or failure of downstream experiments.

The technical solution of the present application will be clearly and completely described below with reference to the accompanying drawings.

Fig. 1 is a schematic structural view of a gene sequencing pretreatment apparatus 1 according to an embodiment of the present application. The gene sequencing pretreatment apparatus 1 includes: the system comprises a rack 100, a kit 200, a pipettor 300, a main control module 800, a station switching module 400, a temperature control module 500 and a magnetic suction module 600; the reagent box 200 is arranged in the rack 100, and a plurality of functional hole sites 210 are arranged on the reagent box 200; the pipettor 300 is disposed in the rack 100; the station switching module 400 is disposed in the rack 100, and is configured to drive the pipettor 300 and/or the reagent kit 200, so that the pipettor 300 is transferred between the functional hole sites 210; the temperature control module 500 is disposed on the rack 100, the reagent cartridge 200 and/or the pipette 300, and is configured to adjust the temperature; the magnetic module 600 is arranged on the kit 200, one or more magnetic modules 600 are arranged, and one magnetic module 600 is positioned at one functional hole site 210 and used for controlling magnetic attraction; the main control module 800 is electrically connected to the station switching module 400, the temperature control module 500 and the magnetic module 600.

The main control module 800 includes a built-in electrical cabinet disposed in the rack 100, and includes: the system comprises a power supply unit, a human-computer interaction interface, a communication unit, a processor and a control unit. The power supply unit can be an external power supply or a storage battery. The man-machine interaction interface can be computer input and output equipment such as a display screen, a keyboard, a touch screen, keys, a knob, a sound box, an LED lamp and the like, and is used for inputting instructions and reading information, so that man-machine interaction and information intercommunication are realized. The communication Unit may be a transceiver, and the control Unit may be a Microcontroller (MCU).

The main control module 800 may receive the instruction and the data through the human-computer interface and transmit the instruction and the data to the processor, and may transmit the message through the human-computer interface to prompt the operator. The main control module 800 receives the temperature information collected by the temperature control module 500 through the communication unit, and transmits the temperature information to the processor. The main control module 800 processes information fed back by the human-computer interaction interface and the communication unit through the processor, and controls the temperature control module 500, the station switching module 400 and the magnetic suction module 600 through the control unit.

Therefore, the working position switching module 400 can be controlled by the main control module 800 so that the liquid transfer device 300 can be transferred among the functional hole sites 210, the magnetic suction module 600 can be controlled to adjust the magnetic suction, and the temperature control module 500 can be controlled to adjust the temperature, so that the integrated functions of nucleic acid extraction, library construction and PCR amplification can be realized, the automation degree is high, the operation difficulty is reduced, and the working efficiency is improved. During the use, only need the kit 200 of dress reagent put into the particular position can, the overall process is inside full-automatic realization in gene sequencing pretreatment device 1, easy operation, relatively with artifical convenient fast, compare with many equipment simple more convenient.

FIG. 2 is an exploded view of a part of the structure of a pretreatment apparatus 1 for gene sequencing according to an embodiment of the present invention. The rack 100 may be assembled by a plurality of supporting sheet metal members, and is used to fixedly mount other components in the gene sequencing pretreatment apparatus 1. The shape of the rack 100 may be designed according to other components in the pretreatment apparatus 1 for gene sequencing. The housing 100 may also be provided with a contamination-resistant enclosure and a clear glass window for viewing.

In this embodiment, the rack 100 is a rectangular parallelepiped structure, and a coordinate system is established with the height direction of the rack 100 as the Z axis, so that the width direction of the rack 100 is the X axis, the length direction of the rack 100 is the Y axis, and the bottom surface of the rack 100 is the X-Y reference plane. Wherein, the X axis, the Z axis and the Y axis are mutually vertical. And the positive direction of the X axis is defined as forward, the positive direction of the Z axis is defined as upward, and the positive direction of the Y axis is defined as rightward.

In order to transfer the pipettes 300 between the functional holes 210, the station switching module 400 has various implementations, for example, the station switching module 400 is a three-dimensional motion system or a two-dimensional motion system, and is connected to the pipettes 300 to move the pipettes 300, while the reagent cartridges 200 are fixed; or, the station switching module 400 is a three-dimensional motion system or a two-dimensional motion system, and is connected with the reagent kit 200 to move the reagent kit 200, while the liquid transfer device 300 is fixed; alternatively, the station switching module 400 is a three-dimensional motion system or a two-dimensional motion system, and is connected to the pipettor 300 and the reagent cartridge 200 at the same time to control the relative movement direction of the pipettor 300 and the reagent cartridge 200.

The third embodiment is adopted, and specifically, the station switching module 400 includes: a first platform 410, an XZ direction movement mechanism 420, a second platform 430 and a Y direction movement mechanism 440, wherein the first platform 410 is connected with the pipette 300; the XZ-direction moving mechanism 420 is connected to the first stage 410 for moving the first stage 410 in the X direction or the Z direction; the second platform 430 is connected with the reagent cartridge 200; the Y-direction moving mechanism 440 is connected to the second stage 430 for moving the second stage 430 in the Y-direction.

With this arrangement, not only can relative three-dimensional movement be achieved between the pipette 300 and the reagent cartridge 200, but also the reagent cartridge 200 can be taken out of the rack 100 by the Y-direction movement mechanism 440.

The XZ-direction movement mechanism 420 and the Y-direction movement mechanism 440 may include one or more of a pulley transmission assembly, a gear transmission assembly, a cam transmission assembly, a rack-and-pinion transmission assembly, a cylinder transmission assembly, a hydraulic cylinder transmission assembly, an electric cylinder transmission assembly, and a motor screw transmission assembly.

In one embodiment, the station switching module 400 further includes: and the displacement sensors are connected with the main control module 800, are respectively arranged on the XZ direction movement mechanism 420 and the Y direction movement mechanism 440, and are used for detecting the displacement of the pipettor 300 and the reagent kit 200.

FIG. 3 is an exploded view of a part of the structure of a pretreatment apparatus 1 for gene sequencing according to an embodiment of the present application. The XZ-direction movement mechanism 420 includes: an X-direction lead screw 421, an X-direction drive motor 422, a Z-direction lead screw 426, and a Z-direction drive motor 427. Both ends of the shaft of the X-direction lead screw 421 are rotatably mounted on the frame 100 by means of a fixing seat or the like; the X-direction driving motor 422 is in transmission connection with the X-direction lead screw 421, and the X-direction driving motor 422 is used for driving the X-direction lead screw 421 to rotate, so that the X-direction driving motor 422 and the connecting member 423 move along the X direction.

Two ends of a shaft of the Z-direction lead screw 426 are rotatably mounted on the connecting piece 423 through fixing seats and the like, and a nut block of the Z-direction lead screw 426 is connected with the first platform 410 and used for driving the first platform 410 to move along the Z direction; the Z-direction driving motor 427 is in transmission connection with the Z-direction lead screw 426 and can drive the Z-direction lead screw 426 to rotate.

To save the installation space, the X-direction driving motor 422 is a through motor. In an operation process, the X-direction driving motor 422 drives the X-direction lead screw 421 to rotate, so that the X-direction driving motor 422 moves back and forth on the X-direction lead screw 421 to drive the connecting member 423 to move back and forth, and the connecting member 423 drives the Z-direction lead screw 426, the Z-direction driving motor 427 and the first platform 410 to move back and forth, thereby realizing the X-direction movement of the liquid dispenser 300 on the first platform 410. The Z-direction driving motor 427 drives the Z-direction lead screw 426 to rotate, so that the first platform 410 moves up and down on the Z-direction lead screw 426, thereby realizing the Z-direction movement of the liquid dispenser 300 on the first platform 410.

The XZ-direction movement mechanism 420 further includes: synchronous belt drive assembly 429, synchronous belt drive assembly 429 include two synchronous pulleys and a synchronous belt, and a synchronous pulley is connected with Z direction driving motor 427, and another synchronous pulley is connected with Z direction lead screw 426. Wherein, Z direction motor and Z direction lead screw 426 locate the both sides of Z direction guide rail 424 respectively, and synchronous belt drive assembly 429 locates the top of Z direction motor and Z direction lead screw 426. With this arrangement, the overall height of the XZ-direction movement mechanism 420 can be reduced, and the occupied space thereof can be reduced.

The XZ-direction movement mechanism 420 further includes: z-direction guide rails 424. The Z-direction guide rail 424 is arranged on the connecting piece 423 along the Z direction, and at least one Z-direction sliding block 425 is arranged on the Z-direction guide rail 424; the Z-direction lead screw 426 is connected to the connecting member 423 via a Z-direction guide rail 424, and a nut block of the Z-direction lead screw 426 is connected to the Z-direction slider 425. With such an arrangement, the Z-direction movement of the first platform 410 is stable, and the first platform is prevented from shifting and vibrating during the movement process.

The XZ-direction movement mechanism 420 further includes: the X-direction guide rail sliding block assembly 4211 and the X-direction guide rail sliding block assembly 4211 comprise two X-direction guide rails which are arranged in parallel, an X-direction sliding block is arranged on one X-direction guide rail, and the X-direction sliding block is connected with the connecting piece 423. With the arrangement, the stress points of the connecting member 423 can be increased, so that the movement of the first platform 410 in the X direction can be stabilized, and the first platform can be prevented from shifting and vibrating during the movement.

The connecting member 423 may include one or more sheet metal members, and the first platform 410 may also include one or more sheet metal members.

The Y-direction movement mechanism 440 includes: the Y-direction guide rail assembly comprises a Y-direction lead screw 443, a Y-direction driving motor 444 and at least one Y-direction guide rail 441, wherein the Y-direction driving motor 444 is in transmission connection with the Y-direction lead screw 443 and can drive the Y-direction lead screw 443 to rotate; both ends of the shaft of the Y-direction screw 443 are mounted on the rack 100 through mounting seats; the nut block of the Y-direction lead screw 443 is connected with the second platform 430, and is used for driving the second platform 430 to move along the Y-direction, the Y-direction guide rail 441 is arranged on the rack 100 along the Y-direction, at least one Y-direction slider 442 is arranged on the Y-direction guide rail 441, and the Y-direction slider 442 is connected with the second platform 430.

In an operation process, the Y-direction driving motor 444 drives the Y-direction lead screw 443 to rotate, so that the second platform 430 moves left and right on the Y-direction lead screw 443, and the Y-direction movement of the reagent cartridge 200 on the second platform 430 is realized.

In this embodiment, two Y-direction guide rails 441 are disposed and respectively located at two sides of the Y-direction lead screw 443, so that the Y-direction movement of the second platform 430 is stable, and the second platform is prevented from shifting and vibrating during the moving process.

The second stage 430 includes: a first substrate 431 and a second substrate 432, the first substrate 431 being connected to the cartridge 200; the second substrate 432 is connected to the first substrate 431 by a plurality of connection bars 436, and is disposed in parallel with the first substrate 431. In this embodiment, there are 4 connecting rods 436.

With this arrangement, an equal space for placing the suction head 920 and the temperature control member can be left between the first base plate 431 and the second base plate 432. The second substrate 432 may further be provided with a diversion hole 433, so as to realize air diversion in the rack 100 and further adjust the detection environment in the rack 100.

The plurality of functional holes 210 includes: at least one tip storage well 216, at least one thermal cover storage well 211, at least one tube storage well 212, at least one first reaction well 213, at least one second reaction well 214, and a plurality of reagent wells 215.

The type, number and arrangement of the plurality of functional holes 210 can be designed according to the requirement, in this embodiment, there are 3 tip storage holes 216 for storing the tips 920 and the tips 920; the test tube storage holes 212 are provided two for respectively storing one test tube with a cap, and the hot cap storage holes 211 are provided one for storing one hot cap 910. One first reaction well 213 is provided and is a micro-volume reaction well. One of the second reaction wells 214 is a large-capacity reaction well. The reagent holes 215 are 28 and distributed in an array. Reagent hole 215 includes circular port and quad slit two kinds, can divide into ordinary reagent district, waste liquid district, trace reagent district and cold-stored reagent district as required. Therefore, the gene sequencing pretreatment device 1 of the embodiment only has one kit 200 for work, is suitable for single sample detection, has the advantages of quick speed and capability of avoiding mutual pollution when a single sample is detected, and can realize most of works in the early stage of gene sequencing, such as nucleic acid extraction, DNA fragmentation, chain length screening, gene purification, DNA/RNA terminal completion, DNA/RNA joint addition, PCR amplification reaction and the like.

In this embodiment, the reagent container 200 is hinged with a flip 217 having an opening, so that the reagent container 200 has an open structure. When the hole-containing flip 217 is closed, the plurality of functional hole sites 210 are exposed through the holes on the hole-containing flip 217. By such arrangement, when the liquid transfer device 300 or the heating column 511 performs operations such as liquid taking, suction head 920 taking, hot cover 910 taking on the kit 200, the flip cover 217 with the hole does not need to be frequently opened, the operation can be directly performed, the procedures are saved, the working efficiency is high, the application range is wide, and the cost is reduced.

FIG. 4 is a sectional view of a pretreatment apparatus 1 for gene sequencing according to an embodiment of the present application. The gene sequencing pretreatment apparatus 1 further includes: the environmental control module 700, the environmental control module 700 is disposed in the rack 100 and connected to the main control module 800, for adjusting an environmental condition in the rack 100.

Therefore, the environmental control module 700 can adjust the environmental conditions inside the rack 100 to form an environment suitable for the operation of the reagent cartridge 200 with an open structure, so as to avoid contamination between samples.

The environment control module 700 includes: at least one ultraviolet light source 710, at least one filter 720 and a ventilation mechanism 730, wherein the ultraviolet light source 710 is disposed on the top of the housing 100 for sterilization and contamination prevention. A plurality of vents 110 are provided in the rack 100. The rack 100 is fully enclosed except for the vents 110.

A filter 720 is provided at each of the ventilation openings 110 for filtering the medical waste gas to prevent external contamination and contamination to the outside. The Filter 720 may be a High efficiency air Filter (HEPA). The ventilation mechanism 730 is disposed within the housing 100 and cooperates with the plurality of ventilation openings 110 to control the wind pressure within the housing 100. The ventilation mechanism 730 includes at least one air inlet 731 and at least one air outlet 732.

In this embodiment, the air inlet parts 731 and the air outlet parts 732 are fans, one air inlet part 731 is provided, and two air outlet parts 732 are provided, wherein the air volume of the air inlet part 731 is smaller than the sum of the air volumes of the two air outlet parts 732, and the air volume of one air inlet part 731 is larger than the air volume of any one air outlet part 732. So set up, can reduce the rotational speed control requirement to single fan, only need through the switch of control fan can.

Specifically, the air inlet 731 is disposed on the bottom surface of the rack 100, and is used for blowing air into the rack 100. An air outlet 732 is disposed at the bottom of the right sidewall of the rack 100 and below the second base plate 432 for exhausting air to the outside of the rack 100. An air outlet 732 is disposed at the top of the rack 100 for exhausting air to the outside of the rack 100. The branch flow port may also be used for air flow communication between the air inlet 731 and the top air outlet 732.

In an operation process, if the air inlet piece 731 and the air outlet piece 732 arranged at the bottom of the rack 100 are opened, the air inlet amount of the rack 100 is larger than the air displacement, the interior of the gene sequencing pretreatment device 1 is positive pressure, so that an external pollution source can be prevented from entering the interior of the gene sequencing pretreatment device 1, and the air volume in the gene sequencing pretreatment device 1 is mainly used for radiating heat of the second table top and the kit 200 on the second table top.

If the air inlet piece 731 and the air outlet piece 732 arranged at the top of the rack 100 are opened, the air inflow of the rack 100 is larger than the air displacement at the moment, the interior of the gene sequencing pretreatment device 1 is positive pressure, so that an external pollution source can be prevented from entering the interior of the gene sequencing pretreatment device 1, and the air volume in the gene sequencing pretreatment device 1 is mainly used for purifying the air in the device.

If the air inlet part 731 and the two air outlet parts 732 are opened simultaneously, the air input of the rack 100 is smaller than the air output, and the inside of the pretreatment device 1 for gene sequencing is negative pressure, so that the pollution source inside the device can be prevented from entering the outside.

In summary, the environmental control module 700 can adopt a positive-negative pressure conversion mode, and is applicable to various samples, and when a sample with strong pollution, such as new coronavirus, is used, the negative pressure in the device is adopted to prevent the sample from polluting the outside; when macro-gene detection is performed on severe patients, external microorganisms are prevented from polluting samples, positive pressure inside the device can be adopted, two air channels for heat dissipation and bacterium removal are arranged inside the device, and the applicability is higher. Therefore, the rack 100 is totally enclosed except the vent 110, and in this embodiment, the ventilation mechanism 730 disposed at the vent 110 cooperates with the ultraviolet light source 710 to sterilize in the environment control module 700, so that a sealed and pollution-free environment is formed inside the gene sequencing pretreatment apparatus 1.

In another embodiment, the ventilation mechanism 730 further comprises at least one wind pressure monitor for detecting wind pressure in the rack 100.

Fig. 5 is a schematic diagram showing a partial structure of the pretreatment apparatus 1 for gene sequencing according to an embodiment of the present application. Fig. 6 is a schematic diagram of the connection between the pipette tip 311 and the pipette tip 920 according to an embodiment of the present application. The pipette 300 includes: the pipette comprises a pipette cavity 310 and a pipette driving member 320, wherein the pipette cavity 310 is connected with a pipette tip 311, the pipette tip 311 can be matched with a suction head 920, and the suction head 920 is hung on the pipette tip 311 and sealed through tension; the pipetting driving member 320 is connected with the pipetting cavity 310 and is used for controlling pipetting of the pipetting tip 311.

The liquid suction and discharge effect is achieved by controlling the air pressure in the suction head 920 on the pipette tip 311 by the pipette driving member 320. The pipetting driver 320 can be a motor or a cylinder, and the piston moves linearly in the pipetting cavity 310, so that the pipette head 920 can suck and discharge liquid by using the air pressure displacement principle.

The pipetting cavity 310 is arranged on the first platform 410, and the pipetting cavity 310 and the pipetting tip 311 thereof are moved by the XZ direction movement mechanism 420, so that the pipetting tip 311 can be controlled to reach the designated functional hole position 210, and operations such as pipetting, pipette tip 920 and the like can be realized.

The pipette 300 further includes: the pipetting detector 330, pipetting detector 330 can include an air pressure sensor and/or a temperature sensor 501, which is disposed in the pipetting cavity 310 and is used for detecting the air pressure and/or temperature of the pipetting cavity 310.

The pipette 300 further includes: the pipette tip withdrawing mechanism 340, the pipette tip withdrawing mechanism 340 including a pipette tip withdrawing plate 341, the pipette tip withdrawing plate 341 being configured to be movable along the axis of the pipette tip 311. When the tip withdrawing movement plate 341 moves downward along the axis of the pipette tip 311, the tip 920 fitted over the pipette tip 311 can be made to fall off.

In order to prevent the overflow of the aerosol, a thermal cap 910 may be attached to the first reaction well 213 for performing the PCR amplification reaction, and the temperature control module 500 includes: a hot lid heating unit 510 for heating the hot lid 910, the hot lid heating unit 510 being disposed in the rack 100 above the reagent cartridge 200. The hot cover heating unit 510 comprises a heating column 511 and an annealing cover mechanism 512, wherein the heating column 511 comprises a heating device and a heat conducting member, and when the hot cover 910 is sleeved on the heating column 511, the hot cover 910 can be heated; the thermal cover withdrawing mechanism 512 comprises a thermal cover moving plate 5121, the thermal cover moving plate 5121 is configured to move along the axis of the heating column 511, and the thermal cover withdrawing mechanism 341 can make the suction head 920 sleeved on the pipette tip 311 fall off when moving downwards along the axis of the heating column 511.

In this embodiment, the heating column 511, the suction head withdrawing mechanism 340 and the heat lid withdrawing mechanism 512 are all disposed on the first platform 410, and may share one XZ movement mechanism, so as to achieve the synchronous movement of the liquid transfer device 300, the heating column 511, the suction head withdrawing mechanism 340 and the heat lid withdrawing mechanism 512, thereby saving the cost and reducing the volume.

In another embodiment, the heating column 511 is still mobile, and a separate heating column 511 moving mechanism is provided to move the heating column 511; the tip withdrawing mechanism 340 and the tip withdrawing mechanism 512 may be disposed at the heating column 511 and the pipette 300, respectively. In another embodiment, the heating column 511 may be fixed, and the thermal cover 910 may be moved to the heating column 511 by adding a moving mechanism for the thermal cover 910.

In this embodiment, the axis of the heating column 511 is parallel to the axis of the pipette tip 311, the moving plate 341 of the pipette tip is integrally connected to the moving plate 5121 of the thermal head, and both the moving plate 341 of the pipette tip and the moving plate 5121 of the thermal head are connected to the lifting/lowering driving assembly 514 in a transmission manner. The lifting driving assembly 514 can be shared to realize the synchronous movement of the suction head removing moving plate 341 and the heat removing cover moving plate 5121, thereby saving the cost and reducing the volume. The connection mode of the moving plate 341 of the suction head and the moving plate 5121 of the heat-removing cover may be integral molding, welding, bolt connection, etc.

In the embodiment, the reagent kit 200, the test tube with the cover, the suction head 920 and the heat cover 910 all adopt disposable consumables, and the suction head 920 or the heat cover 910 can be automatically unloaded through the suction head withdrawing mechanism 340 and the heat cover withdrawing mechanism 512, so that pollution is avoided, human intervention is not required, and the operation is convenient.

The lifting driving assembly 514 may include one or more of a pulley driving assembly, a gear driving assembly, a cam driving assembly, a rack and pinion driving assembly, a cylinder driving assembly, a hydraulic cylinder driving assembly, an electric cylinder driving assembly, and a motor screw driving assembly. For example, the suction head removing moving plate 341 and the heat removing cover moving plate 5121 can be directly driven to lift by an air cylinder or a motor, and the movement of the suction head removing moving plate 341 and the heat removing cover moving plate 5121 can be guided by a guide rod.

Fig. 7 is a schematic structural diagram of a reagent zone temperature control unit 520 according to an embodiment of the present application. Referring to FIG. 8, a cross-sectional view of a reagent zone temperature control unit 520 is shown according to an embodiment of the present application. The temperature control module 500 includes: and the reagent zone temperature control unit 520 is arranged at the bottom of the reagent kit 200, and is used for controlling the temperature of the materials in the partial reagent holes 215 on the reagent kit 200.

In one embodiment, the reagent zone temperature control unit 520 includes: the first heat transfer block 521, the first heat preservation sleeve 522, the first semiconductor refrigeration sheet 523, the first radiator 524 and the first water absorbing piece 526; the first heat transfer block 521 is arranged at the bottom of the kit 200; the first heat-preserving sleeve 522 is sleeved outside the first heat-transferring block 521; the first semiconductor refrigeration sheet 523 is provided with a first cold surface and a first hot surface, and the first cold surface is connected with the first heat transfer block 521; a first heat sink 524 is disposed on the first hot side; the first water absorbing member 526, which may be absorbent cotton, is disposed between the first heat sink 524 and the first heat transfer block 521, and is disposed outside the first semiconductor cooling plate 523.

The first heat sink 524 may include a fin, a vortex fan, and the like. The air inlets 434 of the first heat sink 524 are located on the front and rear sidewalls, and supply air to the front and rear sides, and the air outlets 435 are located at the bottom and supply air to the front. In the embodiment, the region where the reagent holes 215 are located in the lower surface of the reagent kit 200 is cooled by using the vertical temperature difference principle of the semiconductor cooling plate, so that the cooling effect is achieved, and hot air is discharged by the vortex fan of the first radiator 524. The reagent well 215 corresponding to the reagent zone temperature control unit 520 can be used for refrigerating a temperature-sensitive reagent such as an enzyme, etc.

Fig. 9 is a schematic structural diagram of a temperature control unit 530 in a reaction area according to an embodiment of the present disclosure. Referring to fig. 10, a cross-sectional view of a temperature control unit 530 of a reaction region according to an embodiment of the present application is shown. The temperature control module 500 includes a reaction zone temperature control unit 530, and the reaction zone temperature control unit 530 is disposed at the bottom of the kit 200 and is used for controlling the temperature of the materials in the first reaction hole 213 and the second reaction hole 214.

The reaction zone temperature control unit 530 includes: a heating jacket 537 is provided at the bottom of the reagent cartridge 200, and the heating jacket 537 has a first hole 5371 for receiving the second reaction hole 214. The heating sleeve 537 may include heating devices such as annular ceramic heating sheets and heat conducting devices, which are all tubular structures and can avoid interference with the magnetic module 600. The heat dissipation mode of the heating sleeve 537 may be natural heat dissipation without a heat sink.

The reaction zone temperature control unit 530 further includes: the second heat transfer block 531 is arranged at the bottom of the kit 200, and the second heat transfer block 531 is provided with a second hole 5311 for accommodating the first reaction hole 213 and is connected with the heating sleeve 537; a second heat-insulating sleeve 532 sleeved outside the second heat-transfer block 531; the second semiconductor chilling plate 533 has a second cold side and a second hot side, the second cold side being connected to the second heat transfer block 531; the second heat sink 534 is disposed on the second hot side; the second water absorbing member 536, which may be absorbent cotton, is disposed between the second heat sink 534 and the second heat transfer block 531, and is disposed outside the second semiconductor chilling plate 533.

In another embodiment, the temperature control module 500 further comprises: and the liquid transferring temperature control unit is arranged on the liquid transferring device 300 and is used for controlling the temperature of the liquid sucked by the liquid transferring device 300.

The temperature control module 500 includes: the plurality of temperature sensors 501, the plurality of temperature sensors 501 are disposed on the rack 100, the reagent cartridge 200 and/or the pipette 300, and are respectively used for corresponding to the reagent zone temperature control unit 520, the reaction zone temperature control unit 530, the pipette temperature control unit and the hot lid heating unit 510, so as to achieve accurate temperature control. For example: as shown in fig. 10, a temperature sensor 501 is provided at the second heat transfer block 531.

The second heat sink 534 may include fins, vortex fans, and the like. The air inlets 434 of the second heat sink 534 are located on the front and rear side walls and supply air to the front and rear sides, and the air outlets 435 are located at the bottom and supply air to the front.

In the embodiment, by using the principle of the vertical temperature difference of the semiconductor refrigeration sheet, when the first reaction hole 213 is heated or refrigerated, the second semiconductor refrigeration sheet 533 and the second radiator 534 both work to realize the refrigeration effect; during heating, the second semiconductor chilling plate 533 and the second heat sink 534 do not work, and heat is conducted to the upper surface of the chilling plate, so that a heating effect is formed.

Fig. 11 is a schematic diagram showing a partial configuration of a pretreatment apparatus 1 for gene sequencing according to an embodiment of the present application. The second substrate 432 is provided with a diversion hole 433, the first heat sink 524 and the second heat sink 534 are both disposed in the diversion hole 433, wherein the air inlets 434 of the first heat sink 524 and the second heat sink 534 are both located between the first substrate 431 and the second substrate 432, and the air outlets 435 of the first heat sink 524 and the second heat sink 534 are located on one side of the second substrate 432 departing from the first substrate 431.

Because the air inlets 434 of the first heat sink 524 and the second heat sink 534 are both for front and rear side air intake and are located between the first substrate 431 and the second substrate 432, and the air outlet 435 is located below the second substrate 432 and is for forward air outlet, the first substrate 431 and the second substrate 432 are used as a partition plate, thereby preventing hot air from flowing back and reducing the influence on the reagent in the reagent kit 200.

Fig. 12 is a schematic view of a partial structure of a pretreatment apparatus 1 for gene sequencing according to an embodiment of the present application. Each magnetic module 600 includes: the magnetic attraction driving mechanism 630 is used for magnetically attracting the bottom of the functional hole site 210, and the magnetic attraction driving mechanism 630 is connected with the magnetic member 610 and is used for controlling the working state of the magnetic member 610, that is, whether the magnetic member 610 magnetically attracts reagents in the functional hole site 210.

It should be noted that, in the embodiment, one magnetic module 600 is provided to magnetically attract the second reaction hole 214, and a user may also make the plurality of functional hole sites 210 have a magnetic function as required.

The magnetic member 610 may be a permanent magnet or an electromagnet. The magnetic member 610 has a cylindrical, annular or plate shape. That is, the magnetic module 600 has various forms, for example: may be of only one cylindrical shape; or may have only one ring; or may be in the form of only one sheet; it is also possible to have both a cylindrical and an annular shape; it is also possible to have both a cylindrical and a sheet-shaped; it is also possible to have both an annular and a plate-shaped; it is also possible to have both a cylindrical, an annular and a sheet-shaped.

In one embodiment, the magnetic member is a permanent magnet, and the magnetic attraction driving mechanism 630 controls whether the magnetic member 610 attracts the reagent in the functional hole 210 by moving the magnetic member 610 to change the distance between the magnetic member 610 and the reagent kit 200. The magnetic attraction driving mechanism 630 may include one or more of a pulley transmission assembly, a gear transmission assembly, a cam transmission assembly, a rack and pinion transmission assembly, a cylinder transmission assembly, a hydraulic cylinder transmission assembly, an electric cylinder transmission assembly, and a motor screw transmission assembly. For example, a magnetic member 610 may be directly driven to ascend and descend by an air cylinder or a motor.

In another embodiment, the magnetic member 610 is an electromagnet, and the magnetic driving mechanism 630 is a control circuit. The magnetic member 610 is always fixed on the reagent cartridge 200. The magnetic attraction driving mechanism 630 changes whether the magnetic member 610 has magnetism or not by switching on and off, so as to control whether the magnetic member 610 has a magnetic attraction effect on the reagent in the functional hole 210.

Fig. 13 is a partial cross-sectional view of the gene sequencing pretreatment apparatus 1 according to an embodiment of the present application, when the magnetic attraction module 600 is in a loop attraction state. Referring to fig. 14, a partial cross-sectional view of the gene sequencing pretreatment apparatus 1 according to an embodiment of the present application is shown when the magnetic attraction module 600 is in a bottom attraction state. Please refer to fig. 15, which is a schematic structural diagram of a magnetic module 600 in an initial state according to an embodiment of the present application. Please refer to fig. 16, which is a schematic structural diagram of a magnetic module 600 in a ring attraction state according to an embodiment of the present application. Please refer to fig. 17, which is a schematic structural diagram of a magnetic module 600 in a bottom-suction state according to an embodiment of the present application.

The two magnetic members 610 are respectively a cylindrical magnetic member 611 and an annular magnetic member 612, and the cylindrical magnetic member 611 is used for bottom surface magnetic attraction on the bottom of the functional hole 210; the annular magnetic member 612 is sleeved outside the cylindrical magnetic member 611 and is used for performing annular magnetic attraction on the bottom of the functional hole 210; the magnetic attraction driving mechanism 630 is connected to the cylindrical magnetic member 611 and the annular magnetic member 612, and is configured to control the operating states of the cylindrical magnetic member 611 and the annular magnetic member 612.

The magnetic attraction drive mechanism 630 includes: the magnetic-attraction type lifting device comprises a belt wheel assembly 631, a magnetic-attraction driving motor 632, a first lifting plate 633, a second lifting plate 634 and at least one sliding rail 635, wherein the belt wheel assembly 631 comprises a driving belt 6311 and a driving wheel 6312, and the driving belt 6311 is provided with a first shifting block 6313 and a second shifting block 6314 which are opposite to each other; the magnetic attraction driving motor 632 is in transmission connection with the belt wheel assembly 631 through a transmission wheel 6312, and is used for driving the transmission belt 6311 to rotate so as to control the lifting of the first shifting block 6313 and the second shifting block 6314; the first lifting plate 633 is movably arranged on the sliding rail 635 and is connected with the annular magnetic piece 612 and the first shifting block 6313; the second lifting plate 634 is movably disposed on the sliding rail 635, and is connected to the cylindrical magnetic member 611 and the second shifting block 6314. The driving belt 6311 may be a timing belt or a belt. The drive wheel 6312 may be a pulley or a timing wheel.

The magnetic attraction drive mechanism 630 further includes: the limiting assembly 636 comprises a first suction piece 6361 and a second suction piece 6362 which are matched with each other, the first suction piece 6361 is disposed on the second lifting plate 634, and the second suction piece 6362 is disposed on the sliding rail 635. The first attraction member 6361 and the second attraction member 6362 may be a magnet combination or a magnet-metal combination for excluding the interference of the ring-shaped magnetic member 612 with the cylindrical magnetic member 611. In this embodiment, the first attraction 6361 is an auxiliary magnet disposed on the second lifting plate 634, and the second attraction 6362 is a fixed metal plate disposed on the slide rail 635 or a limiting plate with an iron block.

The magnetic attraction drive mechanism 630 further includes: the reset component (not shown in the drawings), which may be an elastic member such as a spring, is sleeved on the slide rail 635 and is used for resetting with the first lifting plate 633 and the second lifting plate 634.

In an operation process, the magnetic module 600 is in an initial state as shown in fig. 15, and the cylindrical magnetic member 611 and the annular magnetic member 612 do not participate in the work and fall below.

When the magnetic module 600 needs to enter the ring attraction state shown in fig. 13 or fig. 16, the magnetic driving motor 632 drives the driving wheel 6312 to rotate clockwise, so as to drive the driving belt 6311 to rotate clockwise, so as to drive the first shifting block 6313 to ascend, the first shifting block 6313 drives the first lifting plate 633 to ascend, the first lifting plate 633 ascends to drive the annular magnetic member 612 to ascend, and due to the attraction between the first attraction member 6361 and the second attraction member 6362 on the second lifting plate 634, the second lifting plate 634 does not ascend under the action of the attraction force between the first attraction member 6361 and the second attraction member 6362, so that the cylindrical magnetic member 63611 does not ascend, and the magnetic module 600 enters the ring attraction state. At this time, the cylindrical magnetic member 611 falls below, and the annular magnetic member 612 rises to penetrate through the heating jacket 537 of the second reaction hole 214 and surround the bottom end of the second reaction hole 214 for a circle.

When the magnetic module 600 needs to enter the bottom attraction state shown in fig. 14 or fig. 17, the magnetic driving motor 632 drives the driving wheel 6312 to rotate counterclockwise, and drives the driving belt 6311 to rotate counterclockwise, so as to drive the second shifting block 6314 to ascend, the second shifting block 6314 drives the first lifting plate 633 and the second lifting plate 634 to ascend, because the second shifting block 6314 directly acts on the second lifting plate 634, the suction effect between the first suction element 6361 and the second suction element 6362 can be overcome, the first lifting plate 633 and the second lifting plate 634 ascend, so as to drive the annular magnetic element 612 and the cylindrical magnetic element 611 disposed thereon to ascend, the cylindrical magnetic element 611 operates, and the bottom attraction state enters the bottom attraction state. At this time, the cylindrical magnetic member 611 is flush with the annular magnetic member 612, and the cylindrical magnetic member 611 contacts the bottom surface of the second reaction well 214.

In this embodiment, the number of the sliding rails 635 is two, the two sliding rails 635 are connected to the first lifting plate 633 and the second lifting plate 634, and the second lifting plate 634 is located below the first lifting plate 633. In another embodiment, there are two slide rails 635, and the first lifting plate 633 and the second lifting plate 634 are respectively disposed on two different slide rails 635. In another embodiment, only 1 sliding rail 635 is provided, and the first lifting plate 633 and the second lifting plate 634 are both provided on the same sliding rail 635.

As shown in fig. 17, the first shifting block 6313 and the second shifting block 6314 can be respectively sleeved on one or two guide rods 639 through their own holes so as to be capable of moving up and down. The guide rod 639 is arranged to guide the first shifting block 6313 and the second shifting block 6314 to move up and down more stably. The guide bar 639, the slide rail 635 and the cylindrical magnetic member 611 are all cylindrical.

Each magnetic module 600 further includes a first fixing plate 637 and a second fixing plate 638, the first fixing plate 637 is used for fixedly connecting the first substrate 431, and the second fixing plate 638 is used for fixing the magnetic driving motor 632. The plurality of guide bars 639 are interposed between the first fixing plate 637 and the second fixing plate 638. The slide rail 635 may also be fixed to the first fixing plate 637.

Please refer to fig. 18, which is a top view of a kit 200 according to an embodiment of the present application. All the functional holes 210 on the reagent kit 200 are arranged more intensively except the tip storing hole 216, and the reagent kit 200 is provided with identification lines for distinguishing the functional holes 210. The indicia lines 1-5 on the left indicate the number of rows and the indicia lines a-C on the top indicate the number of columns of reagent wells 215 in a square. The identification marks a-f on the bottom indicate the number of columns of the reagent wells 215 and the test tube storage wells 212 having a circular shape.

The test tube storage hole 212 may include a5 hole and a b5 hole. The first reaction well 213 is a W1 well and can be used for PCR amplification reaction; the thermal cover storage hole 211 is an S-hole, and the second reaction hole 214 is a W2 hole, and can be used for holding a lysate proteinase K sample.

The functional hole sites 210 requiring refrigeration of the reagent zone temperature control unit 520 may be a2, a3, a4, b2, b3, b4, a5, and b 5.

The A1 hole, the B1 hole, the B2 hole and the C1 hole can be used for placing washing liquid; the hole C2 can be used for placing waste liquid; a2 well can be used for placing fragmentation enzyme reaction liquid; a3 hole can be used for placing the adaptor enzyme reaction liquid; a4 wells can be used for placing PCR adaptor reagents; the hole b2 can be used for placing the end repairing enzyme reaction solution; b3 hole can be used for placing PCR enzyme reaction liquid; c3 hole can be used to place eluent; c4 wells can be used to place fragmented frag bufferi; hole c5 can be used to place a buffer for adding a connector; d3 well can be used to place fragmentation TE buffer; d4 well can be used to place the end repair buffer; e4 well can be used to store screening magnetic beads; f4 well can be used to store purified magnetic beads.

Referring to FIGS. 1 to 18, the following is a method of using the pretreatment apparatus 1 for gene sequencing, comprising the steps of:

step 01: firstly, preparing liquid according to a preset design, prepackaging all reagents, and sending the reagents to a customer; or the customer himself adds the various reagents by manual or other means.

Step 02: the customer adds the sample and proteinase K upon receipt.

Step 03: the reagent cartridge 200 with the reagent placed therein is placed in a predetermined position on the second stage.

Step 04: after the lid 217 including the hole of the kit 200 is closed, the frame 100 is closed and the gene sequencing pretreatment apparatus 1 is started.

Wherein, the steps 01-04 are preparation processes, and then the nucleic acid extraction process is carried out.

Step 05: the Y-direction moving mechanism 440 moves in cooperation with the XZ-direction moving mechanism 420 so that the pipette tip 311 is positioned above the tip storage hole 216 in the reagent cartridge 200.

Step 06: the Z-direction drive motor 427 drives the pipette 300 to move down, and the pipette 300 takes out the tip 920 stored in the tip storage hole 216 and resets it.

Step 07: the Y-direction moving mechanism 440 moves in cooperation with the XZ-direction moving mechanism 420 so that the pipette tip 311 is positioned above the hole W2, and the Z-direction driving motor 427 drives the pipette 300 to move downward so that the pipette tip 920 enters the hole W2. The movements of the Y-direction movement mechanism 440 and the XZ-direction movement mechanism 420 will not be described in detail.

Step 08: the pipettor 300 was controlled to take a portion of the purified magnetic beads from the f4 well to the W2 well.

Step 09: the pipetter 300 blows the liquid and the heating mantle 537 at the bottom of the W2 well works to start the cell lysis DNA process.

Step 10: after a preset time, the heating sleeve 537 stops heating, the magnetic module 600 at the bottom of the hole W2 works (when the liquid is less, the magnetic module 600 enters a bottom suction state, and when the liquid amount is more, the magnetic module 600 enters a side suction or annular state), and stands for a period of time.

Step 11: the pipette 300 removes all the liquid from the W2 well except the magnetic beads.

Step 12: the liquid transfer device 300 removes part of the washing liquid from the hole A1 and the hole B1 to the hole W2, the magnetic suction module 600 at the bottom of the hole W2 stops working, and the liquid transfer device 300 blows and blows reagents in the hole W2 for a period of time.

Step 13: the magnetic module 600 at the bottom of the W2 well works, and after a period of standing, the pipettor 300 removes all the liquid from the W2 well except the magnetic beads to the C2 well (waste region).

Step 14: the process of step 11-step 13 can be repeated one or more times to make the washing more complete. Wherein, the process from the subsequent step 11 to the step 13 is replaced by using the term "washing magnetic beads".

Step 15: pipette 300 removes a portion of the eluate from well c3 into well W2.

Step 16: the magnetic module 600 at the bottom of the W2 well stops working, and the pipette 300 blows the reagent in the W2 well for a period of time.

And step 17: the magnetic module 600 at the bottom of the W2 hole works and stands for a period of time.

Step 18: the pipette 300 removes all liquid except the magnetic beads, half to half of the W1 well to the product O zone (test tube placed in the test tube storage well 212) for sample.

Step 19: the pipettor 300 takes part of the washing liquid from the wells A1 and B1 to the wells W2, the magnetic attraction module 600 at the bottom of the wells W2 stops working, the pipettor 300 blows the reagents in the wells W2 for a period of time, and the pipettor 300 removes all the liquid including the magnetic beads to the wells C2 (waste liquid zone). Wherein the process of the subsequent step 19 is replaced by the term "wash-hole". When this operation is performed on the W2 well, it is referred to as "washing" the W2 well, and when this operation is performed on the other well, it is referred to as "washing" the other well.

Among them, steps 05 to 19 are nucleic acid extraction processes, and the fragmentation process is performed below.

Step 20: the pipetter 300 takes TE buffer, frag buffer II and fragmentation reaction liquid from d3 hole, c4 hole and a2 hole respectively and adds the TE buffer, frag buffer II and fragmentation reaction liquid to W1 hole.

Step 21: the pipette tip 311 is moved to above the tip 920, and the tip retracting mechanism 340 is moved to retract the tip 920 into the tip storing hole 216.

Step 22: the XZ-direction moving mechanism 420 moves in cooperation with the Y-direction moving mechanism 440 to move the heating column 511 on the first platform 410 above the S-hole, the Z-direction driving motor 427 drives the heating column 511 to move downward, and the heating column 511 is taken out to the heat cover 910 in the S-hole.

Step 23: the thermal cover 910 is moved to the position above the W1 hole, the W1 hole is sealed by moving downwards, the heating column 511 is operated to heat the thermal cover 910, a certain temperature is set, the reaction zone temperature control unit 530 at the bottom of the W1 hole starts to operate, and the heating is carried out for a period of time (fragmentation thermal reaction process).

Step 24: the reaction zone temperature control unit 530 stops working, and after a certain period of heat dissipation, the heating column 511 and the thermal cover 910 are driven to move upward by the Z-direction driving motor 427, so that the hole W1 is opened.

Step 25: the thermal cover 910 is moved to above the reagent cartridge 200 hole, the suction head withdrawing mechanism 340 moves, and the thermal cover 910 is withdrawn to the home position.

Among them, steps 20 to 25 are fragmentation processes, and the following goes into a DNA chain length screening process.

Step 26: the pipette tip 311 moves above the tip storage hole 216, and the pipette 300 picks up the tip 920.

Step 27: pipette 300 transfers all liquid in W1 well to W2 well.

Step 28: pipette 300 removes a portion of the screened beads from e4 well to W2 well.

Step 29: pipette 300 blows a W2 hole for a period of time.

Step 30: the magnetic module 600 at the bottom of the W2 hole works and stands for a period of time.

Step 31: the pipette 300 removes all reagents except the magnetic beads to the f5 well for temporary storage.

Step 32: the magnetic module 600 with the W2 hole stops working, and the W1 hole and the W2 hole are washed.

Step 33: pipette 300 retrieves f5 well buffer reagent back to W2 well.

Step 34: pipette 300 removes partially purified magnetic beads from well f4 to well W2.

Step 35: the pipette 300 blows the liquid for a period of time.

Step 36: the magnetic module 600 at the bottom of the W2 hole works and stands for a period of time.

Step 37: pipette 300 pipettes all reagents except magnetic beads from W2 well to C2 well (waste zone).

Step 38: the magnetic module 600 at the bottom of the W2 hole is not in operation.

Step 39: "Wash beads" (can be repeated once).

Step 40: pipette 300 took the eluate from well c3 to well W2.

Step 41: the magnetic module 600 at the bottom of the W2 well is not operated and the pipette 300 blows the reagent for a period of time.

Step 42: the magnetic module 600 at the bottom of the W2 hole works and stands for a period of time.

Step 43: wash f5 wells.

Step 44: pipette 300 removes all liquid except magnetic beads from well W2 to well f 5.

Step 45: the W2 wells were "washed".

Among these, steps 26 to 45 are DNA chain length selection processes, and the end repair process is entered below.

Step 46: pipette 300 took the end repair ERAT buffer and end repair enzyme reaction solution from well d4 and well b2 to well f5, respectively.

Step 47: a terminal repair reaction was performed.

Wherein, the step 46-the step 47 are end repairing processes, and then a joint adding process is carried out.

And 48: pipette 300 removes the ligation Buffer and the ligation reaction solution from well a3 and c5 to well f5, respectively.

Step 49: a linker addition reaction was performed.

Wherein, the steps 48-49 are the process of adding the joint, and then the process of DNA purification is carried out.

Step 50: pipette 300 transfers all reagents from f5 well to W2 well.

Step 51: pipette 300 removes a portion of the magnetic beads from well f4 to well W2.

Step 52: the pipette 300 blows the reagent for a period of time.

Step 53: the magnetic module 600 at the bottom of the W2 hole works and stands for a period of time.

Step 54: pipette 300 removes all extra-magnetic bead reagents from well W2 to well C2 (waste zone).

Step 55: "Wash beads" (can be repeated once).

Step 56: pipette 300 took the eluate from well c3 to well W2.

And 57: the magnetic module 600 at the bottom of the W2 well is not operated and the pipette 300 blows the reagent for a period of time.

Step 58: the magnetic module 600 at the bottom of the W2 hole works and stands for a period of time.

Step 59: pipette 300 removes all reagents except magnetic beads from well W2 to well W1.

Among them, the step 50 to the step 59 are DNA purification processes, and then, a PCR reaction process is performed.

Step 60: pipette 300 removes all reagents except magnetic beads from well W2 to well W1.

Step 61: the pipette 300 took the PCR mixture and the PCR enzyme reaction solution from the b3 well and the a4 well to the W1 well.

Step 62: the pipette 300 moves to the tip deposit hole 216 to withdraw the tip 920.

And step 63: the heating column 511 takes the hot lid 910 and seals the hole to W1.

Step 64: the heating column 511 heats the hot lid 910, and the reaction zone temperature control unit 530 at the bottom of the W1 hole is turned on for cyclic heating.

Step 65: after a period of time, the thermal cap 910 is opened and the thermal cap 910 is withdrawn from the top of the heating column 511 to the top of the S-hole.

And step 66: the pipette 300 reaches the tip access hole 216 to take the tip 920.

Step 67: the liquid resulting from the reaction is transferred to the product O zone (test tube placed in the test tube holding hole 212) by the pipette 300.

Wherein, the steps 46 to 47 are PCR reaction processes.

It should be noted that the features of the embodiments in the present application may be combined with each other without conflict. The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A pretreatment device for gene sequencing, comprising:

a frame;

the kit is arranged in the rack, and a plurality of functional hole sites are arranged on the kit;

the liquid transfer device is arranged in the rack;

the station switching module is arranged on the rack and used for driving the pipettors and/or the reagent boxes so as to enable the pipettors to be transferred among the functional hole sites;

the temperature control module is arranged on the rack, the kit and/or the pipettor and used for adjusting the temperature;

the at least one magnetic suction module is arranged on the kit, is positioned at one functional hole position and is used for controlling magnetic suction; and

the main control module is connected with the station switching module, the temperature control module and the magnetic suction module.

2. The pre-processing apparatus for gene sequencing of claim 1, wherein each of the magnetic attraction modules comprises:

the magnetic part is used for magnetically attracting the bottom of the functional hole site, and the shape of the magnetic part is cylindrical, annular or sheet-shaped; and

and the magnetic suction driving mechanism is connected with the magnetic part and is used for controlling the working state of the magnetic part.

3. The pre-processing apparatus for gene sequencing of claim 2, wherein in each of the magnetic attraction modules, there are two magnetic parts, namely a cylindrical magnetic part and an annular magnetic part, and the cylindrical magnetic part is used for bottom surface magnetic attraction of the bottom of the functional hole site; the annular magnetic part is sleeved outside the cylindrical magnetic part and used for carrying out annular magnetic attraction on the bottom of the functional hole site.

4. The pre-processing apparatus for gene sequencing of claim 3, wherein the magnetic member is a permanent magnet, and the magnetic driving mechanism comprises:

the belt wheel assembly comprises a transmission belt, and a first shifting block and a second shifting block which are opposite to each other are arranged on the transmission belt;

the magnetic attraction driving motor is in transmission connection with the belt wheel assembly and is used for driving the transmission belt to rotate so as to control the first shifting block and the second shifting block to lift;

at least one slide rail;

the first lifting plate is movably arranged on the sliding rail and is connected with the annular magnetic part and the first shifting block;

the second lifting plate is movably arranged on the sliding rail and is connected with the cylindrical magnetic part and the second shifting block;

the limiting assembly comprises a first suction piece and a second suction piece which are matched with each other, the first suction piece is arranged on the second lifting plate, and the second suction piece is arranged on the sliding rail; and

the resetting component is arranged on the sliding rail and used for resetting the first lifting plate and the second lifting plate.

5. The pre-processing apparatus for gene sequencing of claim 2, wherein the magnetic member is an electromagnet, and the magnetic driving mechanism is a control circuit.

6. The pretreatment device for gene sequencing according to claim 1, further comprising:

and the environment control module is arranged in the rack, is connected with the main control module and is used for adjusting the environment condition in the rack.

7. The pre-processing apparatus for gene sequencing according to claim 6, wherein the environment control module comprises:

at least one ultraviolet light source arranged on the frame;

the filter piece is arranged on the ventilation opening; and

and the ventilation mechanism is arranged in the rack and is matched with the plurality of ventilation openings to control the wind pressure in the rack.

8. The pre-processing apparatus for gene sequencing of any one of claims 1 to 7, wherein the station switching module comprises:

a first platform connected with the pipettor;

the XZ direction moving mechanism is connected with the first platform and is used for enabling the first platform to move along the X direction or the Z direction; and

a second platform connected to the kit;

the Y-direction movement mechanism is connected with the second platform and is used for enabling the second platform to move along the Y direction;

wherein, the X direction, the Z direction and the Y direction are mutually vertical.

9. The pre-processing apparatus for gene sequencing according to claim 8, wherein the XZ-direction movement mechanism includes:

the X-direction lead screw is arranged on the rack along the X direction;

the X-direction driving motor is in transmission connection with the X-direction lead screw and is connected with a connecting piece, and the X-direction driving motor is used for driving the X-direction lead screw to rotate so as to enable the X-direction driving motor and the connecting piece to move along the X direction;

the Z-direction lead screw is arranged on the connecting piece along the Z direction, is in transmission connection with the first platform and is used for driving the first platform to move along the Z direction; and

and the Z-direction driving motor is in transmission connection with the Z-direction lead screw and can drive the Z-direction lead screw to rotate.

10. The pre-processing apparatus for gene sequencing according to claim 8, wherein the Y-direction movement mechanism includes:

the Y-direction lead screw is arranged on the rack along the Y direction, is in transmission connection with the second platform and is used for driving the second platform to move along the Y direction;

the Y-direction driving motor is in transmission connection with the Y-direction lead screw and can drive the Y-direction lead screw to rotate; and

and the Y-direction guide rail is arranged on the rack along the Y direction, and is provided with at least one Y-direction sliding block which is connected with the second platform.

11. The pre-gene sequencing processing apparatus according to claim 8, wherein the pipette comprises:

the liquid transferring cavity is connected with a liquid transferring gun head;

the liquid transferring driving piece is connected with the liquid transferring cavity and is used for controlling liquid transferring of the liquid transferring gun head;

the liquid transfer detection piece is arranged in the liquid transfer cavity and is used for detecting the air pressure and/or the temperature of the liquid transfer cavity; and

and the pipette tip withdrawing mechanism comprises a pipette tip withdrawing moving plate which is configured to move along the axis of the pipette tip.

12. The pre-processing apparatus for gene sequencing according to claim 11, wherein the temperature control module comprises:

the hot cover heating unit is arranged in the rack, is positioned above the reagent kit and is used for heating the hot cover;

the hot cover heating unit comprises a heating column and a heat-removing cover mechanism, and the heating column is used for heating the hot cover; the thermal cover mechanism includes a thermal cover moving plate configured to be movable along an axis of the heating column.

13. The pre-processing apparatus for gene sequencing of claim 12, wherein the heating column, the de-chuck mechanism and the de-chuck mechanism are all disposed on the first platform.

14. The pre-processing apparatus for gene sequencing of claim 13, wherein the tip-removing motion plate is connected to the heat-removing cover motion plate, and both the tip-removing motion plate and the heat-removing cover motion plate are in transmission connection with a lifting driving assembly.

15. The pre-processing apparatus for gene sequencing of claim 8, wherein the plurality of functional wells comprise: at least one tip access well, at least one thermal cover access well, at least one tube access well, at least one first reaction well, at least one second reaction well, and a plurality of reagent wells.

16. The pre-processing apparatus for gene sequencing of claim 15, wherein the temperature control module comprises:

the reagent zone temperature control unit is arranged at the bottom of the reagent kit and is used for controlling the temperature of materials in the reagent holes; and

and the reaction area temperature control unit is arranged at the bottom of the kit and is used for controlling the temperature of the materials in the first reaction hole and the second reaction hole.

17. The pre-processing apparatus for gene sequencing of claim 16, wherein the reagent zone temperature control unit comprises:

the first heat transfer block is arranged at the bottom of the kit;

the first heat-preserving sleeve is sleeved outside the first heat-transfer block;

the first semiconductor refrigeration piece is provided with a first cold surface and a first hot surface, and the first cold surface is connected with the first heat transfer block;

the first radiator is arranged on the first hot surface; and

and the first water absorbing piece is arranged between the first heat transfer block and the first radiator and is arranged outside the first semiconductor refrigeration piece.

18. The pre-processing apparatus for gene sequencing of claim 17, wherein the reaction zone temperature control unit comprises:

a heating jacket disposed at the bottom of the reagent cartridge, the heating jacket having a first well for receiving the second reaction well;

the second heat transfer block is arranged at the bottom of the kit and connected with the heating sleeve, and a second hole for accommodating the first reaction hole is formed in the second heat transfer block;

the second heat-insulating sleeve is sleeved outside the second heat-transfer block;

the second semiconductor refrigerating piece is provided with a second cold surface and a second hot surface, and the second cold surface is connected with the second heat transfer block;

the second radiator is arranged on the second hot surface; and

and the second water absorbing piece is arranged between the second radiator and the second heat transfer block and is arranged outside the second semiconductor refrigerating piece.

19. The pre-processing apparatus for gene sequencing according to claim 18, wherein the second platform includes:

a first substrate connected to the bottom of the reagent cartridge; and

the second substrate is connected with the first substrate through a plurality of connecting rods and is arranged in parallel with the first substrate;

the second substrate is provided with a shunting hole, the first radiator and the second radiator are arranged in the shunting hole, wherein the air inlet of the first radiator and the air inlet of the second radiator are both positioned between the first substrate and the second substrate, and the air outlet of the first radiator and the air outlet of the second radiator are positioned on one side of the second substrate, which deviates from the first substrate.

20. The pre-processing device for gene sequencing of claim 1, wherein the kit is hinged with a flip cover containing a hole.