Multi-reaction device for target NGS totally-enclosed efficient automatic library building

Abstract

The utility model discloses a multi-reaction device for targeted NGS totally-closed efficient automatic library building, which comprises a base, reaction holes and reagent storage holes are defined in the base, the reaction holes are used for placing samples to be detected, the reagent storage holes are used for placing reaction reagents, a reagent transfer mechanism is used for sucking the reaction reagents and putting the reaction reagents into the reaction holes, and the reaction holes are used for placing the reaction reagents. The sealing shell covers the base in a sealing mode, and the reagent transferring mechanism is arranged in the sealing shell. A series of operations such as DNA extraction, DNA fragmentation, terminal repair, linker connection and library amplification in the library construction process can be carried out in the reaction hole of the base, so that the automation of library construction is realized, the experiment time is shortened, the labor cost is reduced, and high efficiency is realized; meanwhile, result errors caused by human factors are reduced, experimental results are more accurate, and repeatability of library building operation is achieved; in a sealed environment, the formation of aerosol can be isolated, and the aerosol is prevented from polluting a library amplification product.

Description

Technical Field

[0001] The utility model relates to the technical field of DNA and RNA sequencing devices, in particular to a multi-reaction device for fully enclosed, efficient and automatic library construction of targeted NGS. Background Art

[0002] High-throughput sequencing technology, also known as next-generation sequencing (NGS), is a technology that can sequence large numbers of DNA or RNA sequences quickly and cost-effectively. Compared to traditional Sanger sequencing technology, NGS can sequence millions of DNA fragments simultaneously, greatly improving sequencing efficiency and throughput.

[0003] However, high-throughput sequencing technology is a complex process involving multiple steps, including nucleic acid extraction, library construction, sequencing, and data analysis. Library construction is the most critical factor in determining whether the entire high-throughput sequencing process can be automated. Existing high-throughput sequencing technologies often use semi-automated methods for library construction, which still requires extensive manual work, consuming significant manpower and time. Human factors, such as improper operator operation, can also affect library construction and sequencing results.

[0004] In addition, in library construction experiments, the most likely form of contamination of library amplification products is aerosol contamination; most library construction instruments currently on the market are open, and aerosols can be formed when air and liquid surface rub against each other. Violent shaking of the reaction tube during operation, opening the lid, aspirating samples, and repeated aspiration of samples by contaminated injection guns can all form aerosols and cause contamination.

[0005] In view of this, how to provide a fully automatic library construction device is an urgent problem that needs to be solved by those skilled in the art. Utility Model Content

[0006] The purpose of the utility model is to provide a multi-reaction device for fully enclosed, efficient and automatic library construction of targeted NGS, so as to solve the problems existing in the prior art and realize fully automatic closed library construction.

[0007] To achieve the above objectives, the present invention provides the following solution: The present invention provides a multi-reaction device for fully enclosed, efficient, and automatic library construction of targeted NGS, comprising:

[0008] A base, wherein a reaction hole and a reagent storage hole are defined in the base, the reaction hole is used to place a sample to be detected, and the reagent storage hole is used to place a reaction reagent;

[0009] A reagent transfer mechanism, which is used to absorb the reaction reagent and put the reaction reagent into the reaction well;

[0010] A sealed shell is provided, wherein the sealed shell seals and covers the base, and the reagent transfer mechanism is arranged in the sealed shell.

[0011] Furthermore, the reaction wells include a first reaction well for side magnetic separation, a second reaction well for bottom magnetic separation, and a third reaction well for library amplification reaction; the first and second reaction wells are adjacently arranged and close to one side of the base, and the third reaction well is close to the other side of the base. There are multiple reagent storage wells, and multiple reagent storage wells are arranged between the first / second reaction wells and the third reaction well. The multiple reaction wells can react simultaneously.

[0012] Furthermore, it also includes a waste liquid pool, which is arranged between the third reaction well and the reagent storage well closest to the third reaction well.

[0013] Furthermore, the reagent transfer mechanism includes:

[0014] a threaded rod, the threaded rod being horizontally arranged above the base along the length direction of the base, and the two ends of the threaded rod being rotatably connected to the sealing shell;

[0015] A sampler bracket, the sampler bracket is threadedly connected to the threaded rod, a sampler is provided on the sampler bracket, and a sampling end of the sampler corresponds to the reaction hole / reagent storage hole;

[0016] A pressure plate is horizontally arranged above the sampler bracket along the length direction of the base, and the pressure plate is slidably arranged in the sealed housing along the up and down directions; the lower surface of the pressure plate is in contact with the sampler, and when the threaded rod rotates, the sampler bracket and the sampler move horizontally along the length direction of the base; the sampler bracket is provided with mounting holes through the upper and lower surfaces, the sampler is arranged in the mounting holes, and the outer fixing sleeve of the sampler is provided with a support spring, the bottom end of the support spring is fixedly provided on the sampler bracket;

[0017] A driving module, wherein the driving module can drive the threaded rod to rotate and can drive the pressing plate to move up and down.

[0018] Furthermore, the sampler includes:

[0019] A sampling tube, the sampling tube is arranged on the sampler bracket;

[0020] The syringe pump is fixedly arranged in the sealed shell, and its operating end passes through the sealed shell and extends to the outside. The syringe pump is connected to the sampling tube through a hose. The syringe pump can absorb the air in the sampling tube and squeeze the air in the sampling tube through negative pressure, so that the sampling tube can extract / deliver reaction reagents.

[0021] Furthermore, the sampling tube includes a sampling portion, a buffer portion and a connecting portion connected in sequence from bottom to top, the inner diameter of the sampling portion is smaller than the inner diameter of the buffer portion, and a plurality of grooves are provided on the lower edge of the sampling portion.

[0022] Furthermore, the injection pump comprises:

[0023] A push rod, wherein the syringe pump defines a piston inner tube body, the push rod is disposed in the piston inner tube body, the tail end of the push rod passes through the sealed housing and extends to the outside, and the drive module is capable of driving the push rod to slide along the piston inner tube body;

[0024] A rubber stopper, the rubber stopper being fixedly arranged at the front end of the push rod and adapted to the shape of the inner tube of the piston;

[0025] A spring, one end of which is fixedly connected to the rubber stopper, and the other end is fixedly connected to the inner side wall of the end face of the piston inner tube body. A through hole is opened on the end face of the piston inner tube body, one end of the through hole is connected to the piston inner tube body, and the other end is connected to the hose.

[0026] Furthermore, the sealed housing is provided with a round cover and a sliding cover.

[0027] Furthermore, the shell includes a first shell and a second shell, the first shell and the second shell are connected by mortise and tenon joints, and the base is provided with connecting holes corresponding to the first shell and the second shell, and the first shell and the second shell can be snapped into the connecting holes.

[0028] Furthermore, it also includes a reaction control device, which has a horizontal and / or lifting mechanism, a heating mechanism and / or an ultrasonic mechanism; the lifting mechanism is arranged below the base, and the heating mechanism and the ultrasonic mechanism are arranged on the lifting mechanism and correspond to the position of the reaction hole. The horizontal and / or lifting mechanism can drive the heating mechanism and the ultrasonic mechanism to rise and / or move horizontally and contact the base.

[0029] Furthermore, the horizontal and / or lifting mechanism includes a motor, a lead screw or a belt motion module.

[0030] The utility model discloses the following technical effects:

[0031] 1. The utility model uses a base to load samples to be tested and various reaction reagents. The reaction reagents are extracted and placed into reaction wells under sealed conditions through a reagent transfer mechanism. Through a matching reaction control system, a series of operations such as DNA extraction, DNA fragmentation, end repair, linker ligation, and library amplification in the library construction process can be performed in the reaction wells of the base, thereby fully automating library construction, shortening experimental time and labor costs, and being highly efficient. At the same time, it reduces errors in results caused by human factors, makes experimental results more accurate, and achieves repeatability of library construction operations. In a sealed environment, it can isolate the formation of aerosols and prevent aerosol contamination of library amplification products.

[0032] 2. The sampling tube includes a sampling part, a buffer part and a connecting part which are connected in sequence from bottom to top. The inner diameter of the sampling part is smaller than that of the buffer part, and a plurality of grooves are provided at the lower edge of the sampling part. On the one hand, the buffer part can be used to increase the sampleable reagent capacity of the sampling tube. According to different library construction requirements, it can adsorb small-volume and large-volume (microliter to milliliter level) reagents, thereby improving the applicability of the device. On the other hand, a plurality of grooves are provided at the lower edge of the sampling part, which can increase the suction cross-sectional area of ​​the sampling part, avoid the liquid from being sucked up, and improve the accuracy of the library construction operation and the accuracy of the reagent delivery.

[0033] 3. The present invention makes a special arrangement of the positions of the first reaction hole, the second reaction hole, the third reaction hole and the reagent storage hole on the base, which can improve the effective stroke of the reagent transfer mechanism and the compactness of the overall structure. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0035] Figure 1 This is a schematic diagram of the structure of the utility model;

[0036] Figure 2 Schematic diagram of the internal structure of the sealed housing;

[0037] Figure 3 Schematic diagram of the threaded rod structure;

[0038] Figure 4 Schematic diagram of the syringe pump structure;

[0039] Figure 5 Schematic diagram of the sampler structure;

[0040] Figure 6Arrange isometric diagrams for reaction wells and reagent storage wells

[0041] Figure 7 Arrange the bottom view for the reaction wells and reagent storage wells

[0042] Figure 8 Arrange the front view for the reaction wells and reagent storage wells;

[0043] Among them, 1. First shell; 2. Second shell; 3. Round cover; 4. Sliding cover; 5. Base; 6. Bottom cover; 7. Threaded rod; 8. Syringe pump; 9. Sampler; 10. Sampler bracket; 11. Press plate; 12. Hose; 13. Support spring; 14. Rod cap; 15. Push rod; 16. Rubber stopper; 17. Spring; 18. First reaction hole; 19. Second reaction hole; 20. Reagent storage hole; 21. Waste liquid tank; 22. Third reaction hole. DETAILED DESCRIPTION

[0044] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0045] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the present invention is further described in detail below with reference to the accompanying drawings and specific implementation methods.

[0046] An embodiment of the present utility model provides a multi-reaction device for fully enclosed, efficient and automatic library construction of targeted NGS, comprising: a base 5, wherein reaction holes and reagent storage holes 20 are defined in the base 5, the reaction holes are used to place samples to be detected, and the reagent storage holes 20 are used to place reaction reagents, the upper surface of the base 5 is covered with a bottom cover 6, and the bottom cover 6 is provided with through holes corresponding to each reaction hole and reagent storage hole 20, the base 5 and the bottom cover 6 are connected by a mortise and tenon structure, and an aluminum film-sealed reagent storage hole 20 is provided between the base 5 and the bottom cover 6, and the sampler 9 can pierce the film to complete the sampling during sampling; a reagent transfer mechanism, the reagent transfer mechanism is used to absorb the reaction reagent and put the reaction reagent into the reaction hole; a sealed shell, the sealed shell is sealed and covers the base 5, and the reagent transfer mechanism is arranged in the sealed shell.

[0047] In this embodiment, the reagent transfer mechanism includes: a threaded rod 7, which is horizontally arranged above the base 5 along the length direction of the base 5. The tooth profile of the threaded rod 7 is set to be trapezoidal and has sufficient wear resistance. The two ends of the threaded rod 7 are rotatably connected to the sealed housing, wherein one end of the threaded rod 7 is installed on the left side of the sealed housing through a bearing assembly, and the other end is connected to the rod cap 14. The threaded rod 7 and the rod cap 14 are fixed by a fastening screw. An O-ring is provided on the rod cap 14. The rod cap 14 passes through the sealed housing and extends to the outside. The sealing ring ensures a sealed internal environment. The sampler bracket 10 is threadedly connected to the threaded rod 7. The sampler 9 is provided on the sampler bracket 10, and the sampling end of the sampler 9 corresponds to the reaction hole / reagent storage hole 20. The pressure plate 11 is horizontally arranged above the sampler bracket 10 along the length of the base 5. The pressure plate 11 is slidably arranged in the vertical direction within the sealed housing. The lower surface of the pressure plate 11 contacts the sampler 9, preventing the sampler 9 from being dislodged from the sampler bracket 10 and also preventing the sampler 9 from rotating. When the threaded rod 7 rotates, the sampler bracket 10 and the sampler 9 move horizontally along the length of the base 5. The sampler bracket 10 is provided with mounting holes on its upper and lower surfaces. The sampler 9 is mounted in the mounting holes. The sampler 9 is fixedly sleeved with a support spring 13. The bottom end of the support spring 13 is fixedly mounted on the sampler bracket 10. The upper edge of the sampler 9 is fixedly connected to the upper area of ​​the support spring 13. The connection area between the two does not affect the compression and expansion functions of the support spring 13. When the pressure plate 11 is pressed down, the spring is compressed, and the sampler 9 moves down. When the pressure plate 11 moves up, the support spring 13 is expanded, and the sampler 9 moves up and resets. In this embodiment, the sampler bracket 10 is provided with a slide rail along the longitudinal direction, and the outer surface of the sampler 9 has a cross-shaped protrusion structure that adapts to the internal structure of the mounting hole. A sliding connector is provided on the right side of the sampler 9. The sliding connector is located outside the mounting hole and is slidably connected to the slide rail. The slide rail structure provides auxiliary support and limit for the sampler 9.

[0048] In this embodiment, the sampler 9 includes: a sampling tube, which is arranged on the sampler bracket 10; a syringe pump 8, which is fixedly arranged in a sealed shell, and its operating end passes through the sealed shell and extends to the outside. The syringe pump 8 is connected to the sampling tube through a hose 12. The syringe pump 8 can absorb the air in the sampling tube and squeeze the air in the sampling tube through negative pressure, so that the sampling tube can extract / put in reaction reagents. The sampling tube includes a sampling part, a buffer part and a connecting part connected in sequence from bottom to top. The inner diameter of the sampling part is smaller than the inner diameter of the buffer part, and the lower edge of the sampling part is provided with multiple grooves. The groove structure is provided on the lower edge of the sampling part, so that the sampler 9 has sufficient liquid absorption cross-sectional area when in contact with the reaction hole or the reagent storage hole 20 to avoid liquid from being sucked up. The wall of the sampler 9 is thin enough and the tip is thin enough. The sampler 9 material is a hydrophobic material or is provided with a hydrophobic coating so that the liquid does not stick to the wall, thereby reducing liquid loss and improving sampling accuracy.

[0049] In this embodiment, the injection pump 8 includes: a push rod 15, a piston inner tube body is defined in the injection pump 8, the push rod 15 is arranged in the piston inner tube body, and its tail end passes through the sealed shell and extends to the outside; a rubber plug 16, the rubber plug 16 is fixedly arranged at the front end of the push rod 15, and the rubber plug 16 is adapted to the shape of the piston inner tube body; a spring 17, one end of the spring 17 is fixedly connected to the rubber plug 16, and the other end is fixedly connected to the inner side wall of the end face of the piston inner tube body, and a through hole is opened on the end face of the piston inner tube body, one end of the through hole is connected to the piston inner tube body, and the other end is connected to the hose 12.

[0050] In this embodiment, a circular cover 3 and a sliding cover 4 are provided on the sealed housing. The circular cover 3 and the sliding cover 4 correspond to the positions of the reaction holes, and the samples to be tested can be taken out after the library is built. The housing includes a first housing 1 and a second housing 2, which are connected by a mortise and tenon structure. The base 5 is provided with connection holes corresponding to the first housing 1 and the second housing 2, and the first housing 1 and the second housing 2 can be snapped into the connection holes. In this embodiment, the circular cover 3 is provided on the second housing 2, and the second housing 2 is provided with a pipetting hole corresponding to the circular cover 3. At the same time, a corresponding pipetting hole is also provided on the pressure plate 11 (the pressure plate 11 has only one pipetting hole, and the other holes are decorative holes). After opening the circular cover 3, the samples to be tested, reagents, etc. can be injected into the reaction holes in the base 5 by a pipette. Before placing the samples to be tested, the sampler 9 and the sampler bracket 10 need to be moved to a position away from the reaction holes.

[0051] This embodiment also includes a reaction control device comprising a lifting mechanism, a heating mechanism, and / or an ultrasonic mechanism. The lifting mechanism is disposed below the base 5, and the heating mechanism and ultrasonic mechanism are disposed on the lifting mechanism and correspond to the positions of the reaction wells. The lifting mechanism can drive the heating mechanism and ultrasonic mechanism to move upward and / or horizontally and contact the base 5. Each reaction well is designed with a thin wall, uniformly heated structure, and a curved bottom surface to facilitate heating by the heating mechanism.

[0052] In this embodiment, the reaction wells include a first reaction well 18 for side magnetic separation, a second reaction well 19 for bottom magnetic separation, and a third reaction well 22 for library amplification reaction; the first reaction well 18 and the second reaction well 19 are arranged adjacent to each other and close to one side of the base 5, and the third reaction well 22 is close to the other side of the base 5. There are multiple reagent storage wells 20 (a total of 14 in this embodiment), and multiple reagent storage wells 20 are arranged between the second reaction well 19 and the third reaction well 22. The waste liquid pool 21 is arranged between the third reaction well 22 and the reagent storage well 20 closest to the third reaction well 22. It should be noted that the first reaction well 18, the second reaction well 19, and the third reaction well 22 can all be heated by a heating mechanism. The first reaction well 18 and the second reaction well 19 can be used for constant temperature heating, and the third reaction well 22 is used for directional heating. The heat generated by it is higher, which is also the reason why the third reaction well 22 is separately arranged on the other side of the base 5. The waste liquid reservoir 21 has a larger volume than the reaction wells and reagent storage wells 20 and is less affected by evaporation. Therefore, the waste liquid reservoir 21 is placed close to the third reaction well 22. The first reaction well 18 and the second reaction well 19 are arranged adjacent to each other, and multiple reagent storage wells 20 are arranged between the second reaction well 19 and the third reaction well 22. This allows the reagent transfer mechanism to increase its effective travel when transferring the sampler 9, saving installation space and improving operational efficiency.

[0053] In this embodiment, a drive module is also included. The drive module is fixedly connected to the tail end of the pressing plate 11, the rod cap 14, and the push rod 15 respectively. The drive module can be a motor that drives the pressing plate 11 to move up and down or rotate (the pressing plate 11 is a curved plate, and its rotation can also achieve a downward pressing function), a rotary motor that drives the rod cap 14 to rotate, a telescopic motor that drives the push rod 15 to move telescopically, a cylinder, etc. The drive module can be connected to a numerical control module such as a PLC to accurately control the speed and number of revolutions of the rod cap 14, thereby achieving the effect of accurately controlling the position of the sampler 9. Of course, the reagent storage hole 20 has a certain inner diameter, so even if the sampler 9 and the reagent storage hole 20 deviate slightly, the reaction reagent extraction operation can be successfully completed. It can accurately control the vertical movement distance of the pressing plate 11 and the sampler 9, and can also accurately control the telescopic distance of the push rod 15, thereby accurately controlling the extraction and delivery amount of the reaction reagent, thereby improving the accuracy of library construction.

[0054] The specific working process is as follows:

[0055] Use a pipette to inject the sample to be tested into the reaction hole in the base 5 (in this embodiment, inject it into the first reaction hole 18), and inject the reaction reagent into each reagent storage hole 20 in turn. Connect the base 5 and the bottom cover 6 with mortise and tenon to fix them, and connect the base 5 with the first shell 1 and the second shell 2 with mortise and tenon to fix them. Cover the round cover 3 and the sliding cover 4, check the sealing, and prepare to start the library building operation after checking that there is no problem.

[0056] The driving module drives the rod cap 14 to rotate and moves the sampler 9 to above the reagent storage hole 20 of the reaction reagent to be used. The driving module drives the pressure plate 11 and the sampler 9 to move downward. During the downward movement of the pressure plate 11, the support spring 13 is compressed, and the sampling tube pierces the film and extends below the liquid level of the reaction reagent. The driving module drives the push rod 15 to stretch the spring 17 and at the same time, the suction hose 12 and the sampling tube form a negative pressure environment, and the reaction reagent is sucked from the reagent storage hole 20 into the sampling tube.

[0057] After the suction is completed, the driving module drives the pressure plate 11 to move upward, and the support spring 13 assists the sampler 9 to move upward and reset. The driving module drives the rod cap 14 to rotate and move the sampler 9 to the first reaction hole 18. The driving module drives the push rod 15 to compress the spring 17 and release the reaction reagent in the sampling tube into the reaction hole.

[0058] When it is necessary to heat the sample to be tested, the same method as above is used, except that the sample to be tested is injected into the third reaction hole 22 in the base 5, and the lifting mechanism is started. The lifting mechanism drives the heating mechanism to move upward, and the heating mechanism is brought into contact with the bottom surface of the third reaction hole 22. The heating mechanism is started to directionally and uniformly heat the third reaction hole 22.

[0059] In the description of the present invention, it should be understood that the terms "longitudinal", "transverse", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc., indicating the orientation or position relationship, are based on the orientation or position relationship shown in the accompanying drawings, and are only for the convenience of describing the present invention, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation. Therefore, they should not be understood as limitations on the present invention.

[0060] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements to the technical solutions of the present invention made by ordinary technicians in this field should fall within the scope of protection determined by the claims of the present invention.

Claims

1. A multi-reaction device for fully enclosed, efficient, and automated targeted NGS library construction, characterized in that: include: A base (5), wherein a reaction hole and a reagent storage hole (20) are defined in the base (5), the reaction hole is used to place a sample to be detected, and the reagent storage hole (20) is used to place a reaction reagent; A reagent transfer mechanism, which is used to absorb the reaction reagent and put the reaction reagent into the reaction well; A sealed shell is provided, wherein the sealed shell is sealed and covers the base (5), and the reagent transfer mechanism is arranged in the sealed shell.

2. A multi-reaction device for fully enclosed, efficient, and automatic library construction for targeted NGS according to claim 1, characterized in that: The reaction wells include a first reaction well (18) for side magnetic separation, a second reaction well (19) for bottom magnetic separation, and a third reaction well (22) for library amplification reaction; the first reaction well (18) and the second reaction well (19) are adjacently arranged and close to one side of the base (5), the third reaction well (22) is close to the other side of the base (5), and there are multiple reagent storage wells (20), and the multiple reagent storage wells (20) are arranged between the first reaction well (18) / the second reaction well (19) and the third reaction well (22).

3. A multi-reaction device for fully enclosed, efficient, and automatic targeted NGS library construction according to claim 2, characterized in that: The invention also includes a waste liquid pool (21), wherein the waste liquid pool (21) is arranged between the third reaction hole (22) and the reagent storage hole (20) closest to the third reaction hole (22).

4. A multi-reaction device for fully enclosed, efficient, and automatic library construction for targeted NGS according to claim 1, characterized in that: The reagent transfer mechanism comprises: a threaded rod (7), the threaded rod (7) being horizontally arranged above the base (5) along the length direction of the base (5), and the two ends of the threaded rod (7) being rotatably connected to the sealing shell; A sampler bracket (10), wherein the sampler bracket (10) is threadedly connected to the threaded rod (7), and a sampler (9) is provided on the sampler bracket (10), wherein the sampling end of the sampler (9) corresponds to the reaction hole / reagent storage hole (20); A pressure plate (11), the pressure plate (11) is horizontally arranged above the sampler bracket (10) along the length direction of the base (5), and the pressure plate (11) is slidably arranged in the sealed shell along the up and down directions; the lower surface of the pressure plate (11) is in contact with the sampler (9), and when the threaded rod (7) rotates, the sampler bracket (10) and the sampler (9) move horizontally along the length direction of the base (5); the sampler bracket (10) is provided with mounting holes through the upper and lower surfaces, and the sampler (9) is arranged in the mounting holes, and the outer fixed sleeve of the sampler (9) is provided with a support spring (13), and the bottom end of the support spring (13) is fixedly arranged on the sampler bracket (10); A driving module is provided, wherein the driving module can drive the threaded rod (7) to rotate and can drive the pressing plate (11) to move up and down.

5. A multi-reaction device for fully enclosed, efficient, and automatic library construction for targeted NGS according to claim 4, characterized in that: The sampler (9) comprises: A sampling tube, the sampling tube being arranged on the sampler bracket (10); A syringe pump (8) is fixedly arranged in the sealed housing, and its operating end penetrates the sealed housing and extends to the outside. The syringe pump (8) is connected to the sampling tube through a hose (12). The syringe pump (8) can absorb the air in the sampling tube and squeeze the air in the sampling tube through negative pressure, so that the sampling tube can extract / put the reaction reagent.

6. A multi-reaction device for fully enclosed, efficient, and automatic library construction for targeted NGS according to claim 5, characterized in that: The sampling tube includes a sampling portion, a buffer portion and a connecting portion connected in sequence from bottom to top. The inner diameter of the sampling portion is smaller than the inner diameter of the buffer portion. The lower edge of the sampling portion is provided with multiple grooves.

7. The multi-reaction device for fully enclosed, efficient, and automatic targeted NGS library construction according to claim 5, characterized in that: The injection pump (8) comprises: A push rod (15), wherein a piston inner tube body is defined in the injection pump (8), the push rod (15) is arranged in the piston inner tube body, and its tail end passes through the sealed housing and extends to the outside, and the drive module can drive the push rod (15) to slide along the piston inner tube body; A rubber stopper (16), the rubber stopper (16) being fixedly arranged at the front end of the push rod (15), and the rubber stopper (16) being adapted to the shape of the inner tube body of the piston; A spring (17), one end of the spring (17) is fixedly connected to the rubber stopper (16), and the other end is fixedly connected to the inner side wall of the end surface of the piston inner tube body, and a through hole is opened on the end surface of the piston inner tube body, one end of the through hole is connected to the piston inner tube body, and the other end is connected to the hose (12).

8. The multi-reaction device for fully enclosed, efficient, and automatic library construction of targeted NGS according to claim 1, characterized in that: The sealed housing is provided with a round cover (3) and a sliding cover (4).

9. The multi-reaction device for fully enclosed, efficient, and automatic library construction of targeted NGS according to claim 1, characterized in that: The housing comprises a first housing (1) and a second housing (2), wherein the first housing (1) and the second housing (2) are connected via mortise and tenon joints, and the base (5) is provided with connection holes corresponding to the first housing (1) and the second housing (2), and the first housing (1) and the second housing (2) can be snapped into the connection holes.

10. A multi-reaction device for fully enclosed, efficient, and automatic targeted NGS library construction according to any one of claims 1 to 9, characterized in that: The invention also includes a reaction control device, which has a horizontal and / or lifting mechanism, a heating mechanism and / or an ultrasonic mechanism; the lifting mechanism is arranged below the base (5), and the heating mechanism and the ultrasonic mechanism are arranged on the lifting mechanism and correspond to the position of the reaction hole; the horizontal and / or lifting mechanism can drive the heating mechanism and the ultrasonic mechanism to rise and / or move horizontally and contact the base (5).

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