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

Abstract

The invention discloses a multi-reaction device for targeted NGS totally-closed efficient automatic library establishment, 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 reagent transfer mechanism is used for transferring the reaction reagents into the reaction holes; 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 present invention relates to the technical field of DNA and RNA sequencing devices, and 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 a large number of DNA or RNA sequences quickly and at low cost in a short period of time. Compared with traditional Sanger technology, NGS technology can sequence millions of DNA fragments at the same time, greatly improving the efficiency and throughput of sequencing.

[0003] However, the high-throughput sequencing technology process is complex, involving multiple links such as nucleic acid extraction, library construction, sequencing, and data analysis. Among them, the library construction link is the most critical factor in determining whether the entire high-throughput sequencing process can achieve automated detection. Existing high-throughput sequencing technologies mostly use semi-automatic methods to construct libraries, and a large amount of manual operation is still required during the library construction process, which consumes a lot of manpower and time. Human factors such as improper operation of operators will also affect the 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 of the library construction instruments on the market are open, and aerosols can be formed when air rubs against the liquid surface. 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 a problem that technical personnel in this field urgently need to solve. Summary of the invention

[0006] The purpose of the present invention is to provide a multi-reaction device for fully closed, 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 object, 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 sealed and covers the base, and the reagent transfer mechanism is arranged in the sealed shell.

[0011] Further, 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 reaction well and the second reaction well are arranged adjacent to each other and close to one side of the base, the third reaction well is close to the other side of the base, and there are multiple reagent storage wells, which are arranged between the first reaction well / the second reaction well 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 hole and the reagent storage hole closest to the third reaction hole.

[0013] Further, the reagent transfer mechanism includes:

[0014] A threaded rod, wherein the threaded rod is horizontally arranged above the base along the length direction of the base, and both ends of the threaded rod are rotatably connected to the sealing shell;

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

[0016] A pressure plate, the 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 shell along the up-down direction; 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 sampler outer fixed sleeve is provided with a support spring, and the bottom end of the support spring is fixedly arranged 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] Further, the sampler comprises:

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

[0020] The injection pump is fixedly arranged in the sealed shell, and its operating end passes through the sealed shell and extends to the outside. The injection pump is connected to the sampling tube through a hose. The injection 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 / put in the reaction reagent.

[0021] Furthermore, the sampling tube includes a sampling portion, a buffer portion and a connecting portion which are 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 at the lower edge of the sampling portion.

[0022] Further, the injection pump comprises:

[0023] A push rod, wherein a piston inner tube body is defined in the injection pump, the push rod is disposed in the piston inner tube body, a tail end of the push rod penetrates the sealed housing and extends to the outside, and the driving module can drive the push rod to slide along the piston inner tube body;

[0024] A rubber plug, which is fixedly arranged at the front end of the push rod and is 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 of which is fixedly connected to the inner side wall of the end surface of the piston inner tube body. A through hole is provided on the end surface of the piston inner tube body, one end of which is connected to the piston inner tube body, and the other end of which 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, 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 inserted 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, and 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] The present invention discloses the following technical effects:

[0030] 1. The present invention loads samples to be detected and various reaction reagents through a base, extracts reaction reagents under sealed conditions through a reagent transfer mechanism and can put them into reaction wells. Through a matching reaction control system, a series of operations such as DNA extraction, DNA fragmentation, end repair, linker connection, and library amplification in the process of library construction can be performed in the reaction wells of the base, so that the library construction is fully automated, the experimental time and labor cost are shortened, and it is highly efficient; at the same time, the error of the results caused by human factors is reduced, the experimental results are more accurate, and the repeatability of the library construction operation is achieved; in a sealed environment, the formation of aerosols can be isolated to avoid aerosol contamination of the library amplification products.

[0031] 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. A plurality of grooves are arranged 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 application scope of the device. On the other hand, a plurality of grooves are arranged at the lower edge of the sampling part, thereby increasing the suction cross-sectional area of ​​the sampling part, preventing the liquid from being sucked up, and improving the accuracy of library construction operation and the accuracy of reagent delivery.

[0032] 3. The present invention makes a special arrangement for 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

[0033] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. 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 creative work.

[0034] Figure 1 It is a schematic diagram of the structure of the present invention;

[0035] Figure 2 It is a schematic diagram of the internal structure of the sealed housing;

[0036] Figure 3 It is a schematic diagram of the threaded rod structure;

[0037] Figure 4 It is a schematic diagram of the structure of the injection pump;

[0038] Figure 5 It is a schematic diagram of the sampler structure;

[0039] Figure 6 Arrange isometric diagrams for reaction wells and reagent storage wells

[0040] Figure 7 Arrange the bottom view for the reaction wells and reagent storage wells

[0041] Figure 8 Arrange the front view for the reaction wells and reagent storage wells;

[0042] Among them, 1. the first shell; 2. the second shell; 3. the round cover; 4. the sliding cover; 5. the base; 6. the bottom cover; 7. the threaded rod; 8. the injection pump; 9. the sampler; 10. the sampler bracket; 11. the pressure plate; 12. the hose; 13. the support spring; 14. the rod cap; 15. the push rod; 16. the rubber plug; 17. the spring; 18. the first reaction hole; 19. the second reaction hole; 20. the reagent storage hole; 21. the waste liquid tank; 22. the third reaction hole. DETAILED DESCRIPTION

[0043] 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 described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0044] 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 embodiments.

[0045] The embodiment of the present invention 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, wherein the reaction holes are used to place samples to be detected, and the reagent storage holes 20 are used to place reaction reagents, wherein the upper surface of the base 5 is covered with a bottom cover 6, wherein the bottom cover 6 is provided with through holes corresponding to the reaction holes and the reagent storage holes 20, wherein the base 5 is connected to the bottom cover 6 by a mortise and tenon structure, wherein an aluminum film-sealed reagent storage hole 20 is provided between the base 5 and the bottom cover 6, wherein a sampler 9 can pierce the film to complete sampling during sampling; a reagent transfer mechanism, wherein the reagent transfer mechanism is used to absorb reaction reagents and place the reaction reagents into the reaction holes; and a sealed shell, wherein the sealed shell is sealed and covers the base 5, and the reagent transfer mechanism is arranged in the sealed shell.

[0046] 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, and the threaded rod 7 is set to a trapezoidal shape, which has sufficient wear resistance. Both 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, and the threaded rod 7 and the rod cap 14 are fixed by a fastening screw, and an O-ring is arranged on the rod cap 14, and the rod cap 14 passes through the sealed housing and extends to the outside, through the O The sealing ring ensures the internal sealing environment; the sampler bracket 10, the sampler bracket 10 is threadedly connected with the threaded rod 7, and the sampler 9 is arranged on the sampler bracket 10, and the sampling end of the sampler 9 corresponds to the reaction hole / reagent storage hole 20; the pressing plate 11, the pressing plate 11 is horizontally arranged above the sampler bracket 10 along the length direction of the base 5, and the pressing plate 11 is slidably arranged in the sealing shell along the up and down direction; the lower surface of the pressing plate 11 contacts the sampler 9, that is, to prevent the sampler 9 from coming out of the sampler bracket 10, and also to prevent the sampler 9 from rotating. 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. The sampler 9 is fixedly sleeved with a support spring 13, and the bottom end of the support spring 13 is fixedly arranged on the sampler bracket 10, and the upper edge of the sampler 9 is fixedly connected to the upper area of ​​the support spring 13, and the connection area between the two does not affect the compression and expansion function 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 in the longitudinal direction, and the outer surface of the sampler 9 has a cross-shaped protruding structure, which is adapted to the internal structure of the mounting hole. A sliding connector is provided on the right side of the sampler 9, which is located outside the mounting hole and is slidably connected to the slide rail, and the sampler 9 is auxiliary supported and limited by the slide rail structure.

[0047] 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 the sealed shell, and its operating end penetrates the sealed shell and extends to the outside, and the syringe pump 8 is connected to the sampling tube through a hose 12, and 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. 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 a plurality of grooves. The groove structure is provided at 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, so as to avoid the liquid from being absorbed, the wall of the sampler 9 is thin enough, the tip is thin enough, and the sampler 9 material is a hydrophobic material or is provided with a hydrophobic coating, so that the liquid does not hang on the wall, thereby reducing liquid loss and improving sampling accuracy.

[0048] 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.

[0049] In this embodiment, a round cover 3 and a sliding cover 4 are provided on the sealed housing, and the round cover 3 and the sliding cover 4 correspond to the positions of each reaction hole, and the sample to be detected can be taken out after the library is built. The housing includes a first housing 1 and a second housing 2, and the first housing 1 and the second housing 2 are connected by a mortise and tenon structure. The base 5 is provided with connecting holes corresponding to the first housing 1 and the second housing 2, and the first housing 1 and the second housing 2 can be inserted into the connecting holes. In this embodiment, the round cover 3 is provided on the second housing 2, and the second housing 2 is provided with a pipetting gun hole corresponding to the round cover 3, and a pipetting gun hole is also provided on the pressing plate 11 (there is only one pipetting gun hole on the pressing plate 11, and the other holes are decorative holes). After opening the round cover 3, the sample to be detected, the reagent, etc. can be injected into the reaction hole in the base 5 by a pipetting gun. Before placing the sample to be detected, the sampler 9 and the sampler bracket 10 need to be moved to a position away from the reaction hole.

[0050] In this embodiment, a reaction control device is also included, and the reaction control device has a 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, and the lifting mechanism can drive the heating mechanism and the ultrasonic mechanism to rise and / or move horizontally and contact the base 5. Each reaction hole is designed to be a thin-walled, uniformly heated structure with an arc-shaped bottom surface, which is convenient for heating by the heating mechanism.

[0051] 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, 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, and the heat generated is relatively high, which is also the reason why the third reaction well 22 is arranged separately on the other side of the base 5. The waste liquid pool 21 has a larger volume than the reaction hole and the reagent storage hole 20 and is less affected by evaporation, so the waste liquid pool 21 is arranged close to the third reaction hole 22. The first reaction hole 18 and the second reaction hole 19 are arranged adjacent to each other, and multiple reagent storage holes 20 are arranged between the second reaction hole 19 and the third reaction hole 22, which can increase the effective stroke of the reagent transfer mechanism when transferring the sampler 9, save installation space, and improve operation efficiency.

[0052] In the present embodiment, a driving module is also included, and the driving module is fixedly connected to the tail end of the pressing plate 11, the rod cap 14 and the push rod 15 respectively. The driving module can be a motor that drives the pressing plate 11 to move up and down or rotate (the pressing plate 11 is an arc-shaped plate, and its rotation can also realize the pressing function), a rotating 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 driving module can be connected to the numerical control module such as PLC for communication, and accurately control the rotation speed and number of rotations of the rod cap 14 to achieve the effect of accurately controlling the position of the sampler 9. Of course, the reagent storage hole 20 has a certain inner diameter, and the extraction operation of the reaction reagent can be successfully completed even if the sampler 9 and the reagent storage hole 20 have a slight deviation. It can accurately control the up and down 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, and improving the library construction accuracy.

[0053] The specific working process is as follows:

[0054] 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), inject the reaction reagent into each reagent storage hole 20 in turn, connect the base 5 with the bottom cover 6 and fix it, connect the base 5 with the first shell 1 and the second shell 2 and fix it with mortise and tenon, 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.

[0055] 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 surface of the reaction reagent. The driving module drives the push rod 15 to stretch the spring 17 and simultaneously suck the hose 12 and the sampling tube to form a negative pressure environment, so that the reaction reagent is sucked from the reagent storage hole 20 into the sampling tube.

[0056] 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 to release the reaction reagent in the sampling tube into the reaction hole.

[0057] When it is necessary to heat the sample to be detected, the same method as above is adopted, except that the sample to be detected 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 heat the third reaction hole 22 in a directionally uniform manner.

[0058] In the description of the present invention, it should be understood that the terms "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside" and "outside" etc., indicating orientations or positional relationships, are based on the orientations or positional relationships 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, and therefore should not be understood as a limitation on the present invention.

[0059] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

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

3. A multi-reaction device for fully enclosed, efficient and automatic library construction for targeted NGS according to claim 2, characterized in that: It also includes a waste liquid pool (21), which 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), wherein the threaded rod (7) is horizontally arranged above the base (5) along the length direction of the base (5), and both ends of the threaded rod (7) are 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 arranged on the sampler bracket (10), and a sampling end of the sampler (9) corresponds to a reaction hole / reagent storage hole (20); A pressure plate (11), wherein 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 housing along the up-down direction; 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 a support spring (13) is provided on the outer fixed sleeve of the sampler (9), and the bottom end of the support spring (13) is fixedly arranged on the sampler bracket (10); A driving module, 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 support (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 in 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 comprises a sampling portion, a buffer portion and a connecting portion which are 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 a plurality of grooves.

7. A multi-reaction device for fully enclosed, efficient and automatic library construction for targeted NGS 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 driving module can drive the push rod (15) to slide along the piston inner tube body; 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 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. 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. A multi-reaction device for fully enclosed, efficient and automatic library construction for 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. A multi-reaction device for fully enclosed, efficient and automatic library construction for targeted NGS according to claim 1, characterized in that: The shell comprises a first shell (1) and a second shell (2), the first shell (1) and the second shell (2) being connected by mortise and tenon joints, the base (5) being provided with connecting holes corresponding to the first shell (1) and the second shell (2), the first shell (1) and the second shell (2) being able to be inserted into the connecting holes.

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