Genetic sequencing reaction apparatus, genetic sequencing system, and genetic sequencing reaction method

Abstract

A gene sequencing reaction apparatus, a gene sequencing system and a gene sequencing reaction method. The gene sequencing reaction apparatus comprises a support platform (8); a soaking container arranged on the support platform (8), the soaking container having a soaking reaction area for containing chemical reagents for a gene sequencing reaction and for soaking a sequencing chip (2) having a DNA sample loading structure on a surface and having loaded a DNA sample in the chemical reagents for a gene sequencing reaction; a temperature control device for controlling the temperature of the chemical reagents in the soaking reaction area; and a transfer device for inserting or withdrawing the sequencing chip (2) from the soaking reaction area. The gene sequencing reaction apparatus can realize a gene sequencing reaction process by soaking a sequencing chip in chemical reagents.

Description

[0001] This application is a divisional application of the patent application with international application number "PCT / CN2017 / 095512", international application date of August 1, 2017, national application number "201780093149.0", entry into the Chinese national phase on January 14, 2020, and invention title "Gene Sequencing Reaction Equipment, Gene Sequencing System and Gene Sequencing Reaction Method". Technical Field

[0002] This invention relates to the field of gene sequencing technology, and in particular to a gene sequencing reaction device, a gene sequencing system, and a gene sequencing reaction method. Background Technology

[0003] With current technology, gene sequencing (also known as DNA sequencing) requires loading DNA sample molecules (sequencing reaction templates) onto a sequencing chip before proceeding. Then, nucleotides are added to the sequencing reaction template to complete the gene sequencing reaction. By detecting the type of added nucleotides, the gene sequence can be determined.

[0004] Chinese utility model patent CN205133580U provides a commonly used sequencing chip. This chip has internal channels where different chemical reagents are injected into the channels from the inlet and discharged from the outlet after flowing through the channels. This process is required for both DNA sample loading and sequencing. This traditional sequencing chip requires the injection of sequencing reagents after the DNA sample is loaded onto the channel surface to induce a sequencing chemical reaction. It has the following drawbacks:

[0005] (1) Chemical reagents can only be used once, resulting in high sequencing costs;

[0006] (2) The flow rate of chemical reagents in the flow channel is uneven, and there are corners that chemical reagents cannot reach. This will lead to uneven and insufficient chemical reaction, which will easily cause gene sequencing errors. Moreover, the flowing chemical reagents can easily wash away the DNA molecules that have been loaded onto the chip surface.

[0007] (3) The pressure generated by chemical reagents in the flow channel can easily cause the chip surface to collapse. Deformation of the chip surface will increase the error rate of DNA sequencing.

[0008] (4) Only the part of the silicon surface inside the flow channel is usable on the chip, and the other parts cannot be effectively utilized, which means that the silicon wafer cannot be utilized to the maximum extent. Summary of the Invention

[0009] The purpose of this invention is to provide a gene sequencing reaction device, a gene sequencing system, and a gene sequencing reaction method that utilizes an immersion method to achieve gene sequencing reaction.

[0010] The first aspect of the present invention provides a gene sequencing reaction apparatus, comprising: a support platform; an immersion container disposed on the support platform, the immersion container having an immersion reaction zone for holding chemical reagents for gene sequencing reaction and for immersing a sequencing chip with a DNA sample loading structure on its surface and already loaded with DNA sample in the chemical reagents for gene sequencing reaction; a temperature control device for controlling the temperature of the chemical reagents in the immersion reaction zone; and a transfer device for inserting the sequencing chip into the immersion reaction zone or removing it from the immersion reaction zone.

[0011] Furthermore, the gene sequencing reaction apparatus includes a plurality of said soaking containers; and / or, said soaking containers include a plurality of said soaking reaction zones isolated from each other.

[0012] Furthermore, the soaking container includes an overflow outlet.

[0013] Furthermore, the temperature control device includes a temperature control unit and a water bath. The water bath is used to hold a liquid that can transfer heat. The soaking container is placed in the water bath. The temperature control unit controls the temperature of the liquid in the water bath to control the temperature of the chemical reagent in the soaking reaction zone.

[0014] Furthermore, the gene sequencing reaction device further includes: an upper storage device for placing the sequencing chip to be subjected to gene sequencing reaction; and / or a lower storage device for placing the sequencing chip after the gene sequencing reaction has been completed.

[0015] Further, the upper part storage device and / or the lower part storage device includes: a storage box, the top of which is open and has a slot and a drain hole, the slot being used to position and place the sequencing chip, and the drain hole being located on the bottom wall of the storage box for draining liquid from the storage box; and a liquid storage box, located below the drain hole for receiving the liquid drained from the drain hole.

[0016] Furthermore, the gene sequencing reaction device also includes a container lid, which is closable and can be placed over the immersion container to prevent the chemical reagents from evaporating.

[0017] Furthermore, the container lid includes a plurality of lids, each of which is configured to correspond to one or more of the soaking reaction zones to prevent the evaporation of chemical reagents in the corresponding soaking reaction zone, and at least one of the lids can be opened and closed independently of the other lids.

[0018] Furthermore, the gene sequencing reaction device also includes a flip-top mechanism, which is driven to open and close the container lid.

[0019] Furthermore, the transfer device includes a connector for connecting to the sequencing chip and a motion mechanism driven by the connector to change the working position of the connector.

[0020] Furthermore, the plurality of soaking reaction zones are arranged in one or more rows laterally; the motion mechanism includes a lateral moving mechanism, a longitudinal moving mechanism, and a vertical moving mechanism, the longitudinal moving mechanism is disposed on the support platform, the lateral moving mechanism is disposed on the longitudinal moving mechanism, the vertical moving mechanism is disposed on the lateral moving mechanism, the connecting part is disposed on the vertical moving mechanism, the longitudinal moving mechanism drives the lateral moving mechanism to move longitudinally, the lateral moving mechanism drives the vertical moving mechanism to move laterally, and the vertical moving mechanism drives the connecting part to move vertically.

[0021] Furthermore, the gene sequencing reaction device also includes a chip holding device, which includes one or more chip mounting positions, on which the sequencing chip is mounted to move the sequencing chip by moving the chip holding device.

[0022] Furthermore, the sequencing chip has the DNA sample loading structure on both sides of its surface; the chip mounting position includes a chip mounting port, and the sequencing chip is mounted in the chip mounting port, which is an open port on both sides.

[0023] Furthermore, the chip holding device includes a chip frame, on which the chip mounting position is provided, the surface of the chip frame is a hydrophobic surface and / or the lower end of the chip frame gradually tapers from top to bottom.

[0024] Furthermore, the chip holding device includes a chip frame with one or more chip mounting positions, and a slot at the upper end of the chip frame; the transfer device includes a gripper for connecting to the chip frame and a motion mechanism drivenly connected to the gripper to change the working position of the gripper, the gripper including a positioning frame and a clamping block, the positioning frame being connected to the motion mechanism, the clamping block being movably disposed on the positioning frame, the positioning frame having a positioning hole for the upper end of the chip frame to be inserted, and the gripper engaging with the slot of the chip frame inserted into the positioning hole to clamp the chip frame.

[0025] Furthermore, the gene sequencing reaction apparatus also includes a control device, wherein the control device is coupled to the temperature control device to control the temperature of the chemical reagent; and / or, the control device is coupled to the transfer device to control the soaking time and / or soaking sequence of the sequencing chip in the soaking reaction zone.

[0026] Furthermore, the gene sequencing reaction device includes the sequencing chip, the surface of which has a DNA sample loading structure.

[0027] Furthermore, the gene sequencing reaction device includes a protective cover, and the immersion container is housed within the protective cover.

[0028] Furthermore, the gene sequencing reaction apparatus includes an air blowing device for blowing away chemical reagents from the surface of the sequencing chip and / or the surface of the chip holder on which the sequencing chip is mounted.

[0029] A second aspect of the present invention provides a gene sequencing system, including a DNA sample loading device and a gene sequencing reaction device, wherein the gene sequencing reaction device is the gene sequencing reaction device according to any one of the first aspects of the present invention.

[0030] A third aspect of the present invention provides a gene sequencing reaction method, the gene sequencing reaction method comprising: adding a chemical reagent for gene sequencing reaction to an immersion reaction zone of an immersion container; controlling the temperature of the chemical reagent in the immersion reaction zone; immersing a sequencing chip having a DNA sample loading structure on its surface and loaded with a DNA sample in the chemical reagent for a predetermined time and then removing it.

[0031] Furthermore, the gene sequencing reaction method includes: adding different chemical reagents for gene sequencing reactions into multiple soaking reaction zones, and sequentially soaking the sequencing chip in the multiple soaking reaction zones for a predetermined time in a predetermined order.

[0032] Based on the gene sequencing reaction apparatus provided by this invention, its immersion container has an immersion reaction zone, which is used to hold chemical reagents for gene sequencing reactions and to immerse a sequencing chip with a DNA sample loading structure on its surface and already loaded with DNA samples in the chemical reagents for gene sequencing reactions; its temperature control device is used to control the temperature of the immersion reaction zone; and its transfer device is used to insert the sequencing chip into or remove it from the immersion reaction zone. Using this gene sequencing reaction apparatus, gene sequencing reactions can be achieved by immersion.

[0033] The sequencing chip is immersed in chemical reagents in different immersion reaction zones, which enables it to complete all the steps required for the sequencing reaction.

[0034] The chemical reagents in the soaking reaction zone can be reused, thereby reducing the cost of consumables.

[0035] The immersion method eliminates the problem of uneven liquid flow rate, and bubbles are less likely to form on the surface of the sequencing chip, ensuring a more uniform and complete chemical reaction.

[0036] The sequencing chip experiences uniform liquid pressure and heating within the immersion tank, thus preventing deformation.

[0037] It has no complex fluid systems, fewer parts, is easy to assemble, and has low manufacturing costs. It can simultaneously soak multiple sequencing chips, offering the advantage of high throughput.

[0038] The gene sequencing reaction equipment is automatically controlled by a control device, enabling automated operation.

[0039] The gene sequencing system and gene sequencing reaction method of the present invention have similar technical effects to the gene sequencing reaction equipment of the present invention.

[0040] Other features and advantages of the invention will become clear from the following detailed description of exemplary embodiments of the invention with reference to the accompanying drawings. Attached Figure Description

[0041] The accompanying drawings, which are included to provide a further understanding of the invention and form part of this application, illustrate exemplary embodiments of the invention and, together with their description, serve to explain the invention and do not constitute an undue limitation thereof. In the drawings:

[0042] Figure 1 This is a three-dimensional structural diagram of the gene sequencing reaction device in an embodiment of the present invention;

[0043] Figure 2 yes Figure 1 A three-dimensional structural diagram of the gene sequencing reaction equipment after the container lid has been removed;

[0044] Figure 3 yes Figure 1 A three-dimensional structural diagram of the gene sequencing reaction device from another angle;

[0045] Figure 4 yes Figure 1 A three-dimensional structural diagram of the transfer device, sequencing chip, and chip holding device of the gene sequencing reaction equipment shown.

[0046] Figure 5 yes Figure 4 A three-dimensional structural diagram of the transfer device's grippers, sequencing chip, and chip holding device from another angle.

[0047] Figure 6 yes Figure 4 The diagram shows an exploded view of the transfer device's grippers, sequencing chip, and chip holding device.

[0048] Figure 7 yes Figure 4 The exploded view of the transfer device's grippers, sequencing chip, and chip holding device from another angle.

[0049] Figure 8 yes Figure 1 A three-dimensional structural diagram of the upper material storage device (lower material storage device) of the gene sequencing reaction equipment shown;

[0050] Figure 9 yes Figure 8 The exploded view of the upper part storage device (lower part storage device) is shown.

[0051] Figure 10 yes Figure 1 A three-dimensional structural diagram of the soaking tank of the gene sequencing reaction equipment shown;

[0052] Figure 11 yes Figure 1 The diagram shows an enlarged view of Part I of the gene sequencing reaction device.

[0053] Figures 1 to 11 In the figures, the labels represent:

[0054] 1. Gene sequencing reaction equipment; 2. Sequencing chip; 3. Immersion tank; 4. Container lid; 5. Temperature control device; 6. Transfer device; 7. Grippers; 8. Support platform; 9. Chip frame; 10. Bayonet; 11. Positioning frame; 12. Clamping block; 13. Positioning hole; 14. Upper part storage device; 15. Lower part storage device; 16. Slot; 17. Drain hole; 18. Storage box; 19. Lateral movement mechanism; 20. Longitudinal movement mechanism; 21. Vertical movement mechanism; 22. Flip-top mechanism; 23. Control device; 24. Chip mounting position; 25. Air blowing device; 26. Push-pull rod; 27. Casters; 28. Indicator light; 29. ​​Overflow port. Detailed Implementation

[0055] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. The following description of at least one exemplary embodiment is merely illustrative and is in no way intended to limit the present invention or its application or use. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0056] Unless otherwise specifically stated, the relative arrangement, numerical expressions, and values ​​of the components and steps set forth in these embodiments do not limit the scope of the invention. It should also be understood that, for ease of description, the dimensions of the various parts shown in the drawings are not drawn to actual scale. Techniques, methods, and devices known to those skilled in the art may not be discussed in detail, but where appropriate, such techniques, methods, and devices should be considered part of the specification. In all examples shown and discussed herein, any specific values ​​should be interpreted as merely exemplary and not as limitations. Therefore, other examples of exemplary embodiments may have different values. It should be noted that similar reference numerals and letters in the following figures denote similar items; therefore, once an item is defined in one figure, it need not be further discussed in subsequent figures.

[0057] In the description of this invention, it should be understood that the use of terms such as "first" and "second" to define components is merely for the purpose of distinguishing the corresponding components. Unless otherwise stated, the above terms have no special meaning and therefore should not be construed as limiting the scope of protection of this invention.

[0058] In the description of this invention, it should be understood that the orientation or positional relationship indicated by directional terms such as "front, back, up, down, left, right", "horizontal, vertical, horizontal" and "top, bottom" is generally based on the orientation or positional relationship shown in the accompanying drawings, and is only for the convenience of describing this invention and simplifying the description. Unless otherwise stated, these directional terms do not indicate or imply that the device or element referred to must have a specific orientation or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation on the scope of protection of this invention; the directional terms "inner" and "outer" refer to the inner and outer contours relative to the outline of each component itself.

[0059] The term "horizontal" as used in the following description refers to Figures 1 to 3 The left and right directions of the gene sequencing reaction device 1 shown, and the "vertical" direction refers to... Figures 1 to 3 The front-to-back direction and "vertical" direction of the gene sequencing reaction device 1 shown are indicated by... Figures 1 to 3 The vertical direction of the gene sequencing reaction device 1 shown.

[0060] Figures 1 to 11 The structure of a gene sequencing reaction apparatus 1 according to an embodiment of the present invention is shown. Figures 1 to 11As shown in the figure, an embodiment of the present invention discloses a gene sequencing reaction apparatus 1. The gene sequencing reaction apparatus 1 includes a support platform 8, an immersion container, a temperature control device 5, and a transfer device 6. The immersion container is disposed on the support platform 8. The immersion container has an immersion reaction zone, which is used to hold chemical reagents for gene sequencing reactions and to immerse a sequencing chip 2, which has a DNA sample loading structure on its surface and is already loaded with DNA samples, in the chemical reagents for gene sequencing reactions. The temperature control device 5 is used to control the temperature of the chemical reagents in the immersion reaction zone. The transfer device 6 is used to insert the sequencing chip 2 into or remove it from the immersion reaction zone.

[0061] In this embodiment, the DNA sample is a sequencing reaction template, and the labeled nucleotides can be added to the sequencing reaction template.

[0062] The gene sequencing reaction device 1 provided by this invention can achieve gene sequencing reactions through an immersion method. The sequencing chip 2 is immersed in chemical reagents in different immersion reaction zones, completing all the steps required for the sequencing reaction. The chemical reagents in the immersion reaction zones can be reused, thereby reducing consumable costs. The immersion method eliminates the problem of uneven liquid flow, and bubbles are less likely to form on the surface of the sequencing chip 2, ensuring a more uniform and complete chemical reaction. The sequencing chip 2 experiences uniform liquid pressure and heating within the immersion reaction zone, thus preventing deformation. It eliminates the need for a complex fluid system, has fewer parts, is easy to assemble, and has low manufacturing costs. Multiple sequencing chips 2 can be immersed simultaneously, offering the advantage of high throughput. The gene sequencing reaction device 1 is automatically controlled by a control device 23, enabling automated operation.

[0063] The following combination Figures 1 to 11 The embodiments of the present invention will be described in detail.

[0064] like Figures 1 to 11 As shown, the gene sequencing reaction device 1 of this embodiment includes a sequencing chip 2, an immersion container, a temperature control device 5, a transfer device 6, a chip holding device, an upper material storage device 14, a lower material storage device 15, a container lid 4, a flip-top mechanism 22, an air blowing device 25, a control device 23, and a support platform 8.

[0065] The sequencing chip 2 has a DNA sample loading structure on its surface. Before the gene sequencing reaction, the DNA sample loading structure of the sequencing chip 2 is already loaded with DNA sample. In this embodiment, the sequencing chip 2 is a silicon wafer, and adapters capable of capturing DNA molecules are pre-set on both sides of the silicon wafer. DNA molecules can be captured by these adapters through a series of chemical reactions and finally adhere to the silicon wafer surface. Adapters can be formed, for example, by modifying the silicon wafer surface with amino groups. During the gene sequencing reaction, the DNA sample remains adhered to the sequencing chip 2.

[0066] The DNA sample referred to in this embodiment can be a nanosphere molecule disclosed in US Patent US8445197B2, also known as DNB. In this process, genomic DNA is first fragmented, then adapter sequences are added, and the DNA is circularized to form single-stranded circular DNA. Subsequently, rolling circle amplification (RoBA) technology is used to amplify the single-stranded circular DNA by two to three orders of magnitude, resulting in DNB.

[0067] The immersion container has an immersion reaction zone, which holds the chemical reagents used in the gene sequencing reaction and is used to immerse the sequencing chip 2, which has a DNA sample loading structure on its surface and is already loaded with DNA, in the chemical reagents for gene sequencing. The sequencing chip 2 is immersed in the chemical reagents in the immersion reaction zone to complete the gene sequencing reaction step. The temperature control device 5 can control the temperature of the chemical reagents in the immersion reaction zone, thereby providing suitable temperature conditions for the gene sequencing reaction.

[0068] In this embodiment, the gene sequencing reaction device 1 includes multiple soaking containers. For example... Figures 1 to 3 As shown, the soaking container is specifically a soaking tank 3. Each soaking tank 3 has a soaking reaction zone. Multiple soaking tanks 3 are arranged on a temperature control device 5. The temperature control device 5 is located on a support platform 8. A transfer device 6 can insert and remove the sequencing chip 2 from the soaking tank 3. In this embodiment, each soaking tank 3 contains a chemical reagent, and different soaking tanks contain the chemical reagents required for each gene sequencing step. After the transfer device 6 holds the sequencing chip 2 and soaks it in each soaking tank 3 for a predetermined time, nucleotides can be added to the DNA sample (sequencing reaction template) of the sequencing chip 2. Then, the sequencing chip 2 is placed on an external optical imaging device for imaging, and the type of added nucleotides can be detected. By continuously repeating the above steps, the gene sequence can be read.

[0069] In other embodiments not shown, the soaking tanks 3 may be grouped, with each group containing a chemical reagent.

[0070] refer to Figures 1 to 3 In this embodiment, the gene sequencing reaction device 1 includes two rows of horizontally arranged soaking tanks 3, with 17 soaking tanks 3 in each row.

[0071] In other embodiments not illustrated, the soaking container may also include multiple soaking reaction zones isolated from each other, each or each group of soaking reaction zones containing a chemical reagent.

[0072] like Figure 10 As shown, in this embodiment, the soaking tank 3 includes an overflow port 29. When the liquid in the soaking tank 3 exceeds a certain water level, it overflows from the overflow port 29, thereby preventing the liquid level in the soaking tank 3 from becoming too high.

[0073] Temperature control device 5 is used to control the temperature of the chemical reagents in the soaking reaction zone. Temperature control device 5 is mounted on support platform 8, and the soaking container is mounted on support platform 8 via temperature control device 5.

[0074] In this embodiment, as Figures 1 to 3 As shown, the temperature control device 5 includes a temperature control unit and a water bath. The water bath is used to hold a liquid capable of transferring heat. The immersion container is placed in the water bath. The temperature control unit controls the temperature of the liquid in the water bath to control the temperature of the chemical reagent in the immersion container.

[0075] In this embodiment, the temperature control unit is a heat exchange tube installed inside the wall of the water bath. The heat exchange tube can input heat into the liquid in the water bath or absorb heat from the liquid in the water bath, so that the heat-conducting liquid in the water bath can be heated or cooled accordingly.

[0076] By inputting or outputting heat to the walls of the water bath or to the liquid inside, the heat-conducting liquid in the water bath can be heated or cooled accordingly. Immersion tank 3 is immersed in the heat-conducting liquid, thus enabling control of the temperature of the chemical reagents within it. Using liquid as the heat transfer medium allows for more uniform temperature control of the chemical reagents in each immersion tank 3. Furthermore, the temperature of the chemical reagents is more stable and less prone to rapid changes.

[0077] In other embodiments not shown, a direct temperature control method can be used to directly heat or cool the soaking tank 3 or the liquid inside it using a temperature controller (e.g., a Peltier temperature controller). However, compared with the indirect temperature control method using a water bath, the direct temperature control method is more prone to uneven heating and cooling.

[0078] The upper storage device 14 is used to place the sequencing chip 2 to be used in the gene sequencing reaction. The lower storage device 15 is used to place the sequencing chip 2 that has already undergone the gene sequencing reaction.

[0079] like Figure 8 and Figure 9 As shown, in this embodiment, both the upper storage device 14 and the lower storage device 15 include a storage box and a liquid storage box 18. The top of the storage box is open and has a slot 16 and a drain hole 17. The slot 16 is disposed on the inner surface of the side wall of the storage box and is used to position and place the sequencing chip 2. The drain hole 17 is disposed on the bottom wall of the storage box and is used to drain the liquid in the storage box. In this embodiment, the drain hole 17 can drain the chemical reagents flowing down the surface of the chip frame 9 and the sequencing chip 2.

[0080] In this embodiment, during the gene sequencing reaction, the sequencing chip 2 is mounted on the chip frame 9, and the sequencing chip 2 is moved by moving the chip frame 9. The slot 16 is used to position and place the chip frame 9, thereby positioning and placing the sequencing chip 2. The chip frame 9 is inserted into the slot 16 to position and place the sequencing chip 2 within the storage container.

[0081] The number of slots 16 in the upper component storage device 14 and the lower component storage device 15 preferably corresponds to the number of positioning holes 13 on the gripper 7, which will be described later, to ensure that the gripper 7 accurately clamps and places the chip frame 9. Figures 4 to 7 As shown, in this embodiment, the number of positioning holes 13 in the gripper 7 is three; as Figure 8 and Figure 9 As shown, the upper part storage device 14 and the lower part storage device 15 each have three slots 16.

[0082] The liquid storage box 18 is located below the drain hole 17 and is used to collect the liquid discharged from the drain hole 17. In this embodiment, the liquid storage box 18 is made into a drawer-type box, which can be removed and emptied when the liquid storage box 18 is full of liquid or when the gene sequencing reaction device 1 is not in use.

[0083] It should be noted that although both the upper storage device 14 and the lower storage device 15 of the gene sequencing reaction device 1 in this embodiment include a storage box and a liquid storage box 18, the specific structures of the upper storage device 14 and the lower storage device 15 can be configured in other ways in other embodiments not shown. The structures of the upper storage device 14 and the lower storage device 15 can be the same or different.

[0084] Furthermore, neither the upper storage device 14 nor the lower storage device 15 is necessary. For example, after the sequencing chip 2 is mounted on the chip holder, it can be directly placed in the soaking reaction zone corresponding to the first reaction step of the gene sequencing reaction without the need for the upper storage device 14. As another example, after the sequencing chip 2 completes the sequencing reaction, it can be directly sent to an external optical imaging device for imaging without the need for the lower storage device 15.

[0085] The container lid 4 is closable and can be placed on the soaking container to prevent the chemical reagents from evaporating.

[0086] like Figure 1 , Figure 2 and Figure 11As shown, preferably, the container lid 4 includes multiple lids, each lid corresponding to one or more soaking reaction zones to prevent the evaporation of chemical reagents within the corresponding soaking reaction zone. At least one lid can be opened and closed independently of the others. In this embodiment, corresponding to two rows of soaking tanks 3, the container lid 4 includes two rows of lids, with six lids in each row. Each lid covers several soaking tanks 3.

[0087] The flip-top mechanism 22 is driven by the container lid 4 to open and close the container lid 4. For example... Figures 1 to 3 As shown, the flip-top mechanism 22 is mounted on the support platform 8. See also... Figure 11 The flip-top mechanism 22 includes a push-pull rod 26 corresponding to the cover body. The push-pull rod 26 is connected to the cover body of the container cover 4. When the push-pull rod 26 pulls the cover body, the cover body opens. When the push-pull rod 26 pushes the cover body, the cover body closes.

[0088] The transfer device 6 is used to insert or remove the sequencing chip 2 from the immersion reaction zone. In this embodiment, the transfer device 6 is used to move the sequencing chip 2 to insert or remove it from each immersion reaction zone.

[0089] The transfer device 6 in this embodiment can move the sequencing chip 2 in one soaking tank 3 to another soaking tank 3.

[0090] In this embodiment, the sequencing chip 2 to be subjected to gene sequencing reaction can be placed into the slot 16 of the upper material storage device 14 by another external transfer device, or the sequencing chip 2 can be placed into the slot 16 of the upper material storage device 14 manually. Similarly, the sequencing chips 2 that have undergone gene sequencing reaction can be removed by another external transfer device, or the sequencing chips 2 can be removed manually.

[0091] The transfer device 6 can improve the automation level of the gene sequencing reaction equipment 1, reduce the error rate caused by manual operation, and can also precisely control the soaking sequence and soaking time by cooperating with the control device 23, thereby facilitating the high-quality completion of the gene sequencing reaction.

[0092] In this embodiment, the transfer device 6 is used to move the sequencing chip 2, and includes a connector for connecting to the sequencing chip 2 and a motion mechanism that is driven to change the working position of the connector.

[0093] The transfer device 6 is mounted on the support platform 8. In other embodiments not shown, the transfer device 6 may also be mounted on other supports, as long as they enable the connection and movement of the sequencing chip 2.

[0094] In this embodiment, as Figures 1 to 3As shown, the motion mechanism includes a lateral moving mechanism 19, a longitudinal moving mechanism 20, and a vertical moving mechanism 21. The longitudinal moving mechanism 20 is mounted on the support platform 8. The lateral moving mechanism 19 is mounted on the longitudinal moving mechanism 20. The vertical moving mechanism 21 is mounted on the lateral moving mechanism 19. A connecting portion is mounted on the vertical moving mechanism 21. The longitudinal moving mechanism 20 drives the lateral moving mechanism 19 to move longitudinally. The lateral moving mechanism 19 drives the vertical moving mechanism 21 to move laterally. The vertical moving mechanism 21 drives the connecting portion to move vertically. In this embodiment, the connecting portion includes grippers 7 for holding the sequencing chip 2.

[0095] As mentioned earlier, multiple soaking tanks 3 are arranged in two rows laterally. By combining the use of the lateral moving mechanism 19, the longitudinal moving mechanism 20, and the vertical moving mechanism 21, it can be ensured that the gripper 7 can insert and remove the sequencing chip 2 from any of the soaking tanks 3.

[0096] Alternatively, the soaking tanks 3 can be arranged in a ring, in which case the motion mechanism can include a rotary mechanism. The connecting part can also be in other forms, such as a vacuum chuck, electromagnetic chuck, etc., that cooperates with the chip holding device for supporting the sequencing chip 2.

[0097] In addition, although in this embodiment the sequencing chip 2 is moved between different soaking tanks 3 by moving the sequencing chip 2 through the motion mechanism and the connecting part, in other embodiments not shown, the desired positional relationship between the sequencing chip 2 and the soaking container can also be achieved simply by moving the soaking container or by moving the soaking container and the sequencing chip 2 at the same time.

[0098] A chip holding device is used to hold the sequencing chip 2 in place, allowing the sequencing chip 2 to move along with the chip holding device. The chip holding device includes one or more chip mounting positions 24, on which the sequencing chip 2 is mounted so as to move the sequencing chip 2 by moving the chip holding device.

[0099] Setting up a chip holding device can reduce contamination caused by direct manipulation of the sequencing chip 2. On the other hand, it can also move multiple sequencing chips 2 simultaneously as needed and maintain a predetermined interval between them, thereby increasing the throughput of the sequencing chip 2.

[0100] In this embodiment, the sequencing chip 2 has DNA sample loading structures on both sides of its surface. For example... Figures 4 to 7 As shown, the chip mounting position 24 includes a chip mounting port, within which the sequencing chip 2 is mounted. The chip mounting port is an open port on both sides. This configuration allows both sides of the sequencing chip 2 to be immersed, increasing the number of DNA sample molecules loaded onto a single sequencing chip 2.

[0101] Furthermore, when transferring the chip frame 9 between different soaking tanks 3, to minimize cross-contamination between different chemical reagents, it is generally necessary to wait until all residual liquid on the surface of the chip frame 9 has dripped off before transferring it to the next soaking tank 3. To accelerate the dripping speed of liquid from the surface of the chip frame 9, such as... Figures 4 to 7 As shown, in this embodiment, the lower end of the chip frame 9 gradually tapers from top to bottom. In an alternative embodiment, the surface of the chip frame 9 can be configured as a hydrophobic surface. Alternatively, the chip frame 9 can have a gradually tapering lower end while also having a hydrophobic surface, thereby enabling faster dripping of chemical reagents.

[0102] like Figures 5 to 7 As shown, a bayonet 10 is provided at the upper end of the chip frame 9; the motion mechanism of the transfer device 6 is driven to change the working position of the gripper 7, and the gripper 7 is used to connect with the chip frame 9.

[0103] In this embodiment, the gripper 7 includes a positioning frame 11 and a clamping block 12. The positioning frame 11 is connected to the motion mechanism, and the clamping block 12 is movably disposed on the positioning frame 11. The positioning frame 11 is provided with a positioning hole 13 for inserting the upper end of the chip frame 9. The gripper 7 engages with the slot 10 of the chip frame 9 inserted into the positioning hole 13 to clamp the chip frame 9.

[0104] like Figures 4 to 7 As shown, each chip frame 9 has two opposing slots 10. The grippers 7 also have two corresponding clamping blocks 12. The two clamping blocks 12 are located at the upper end of the positioning holes 13. When the chip frame 9 passes upward through the positioning holes 13, the two clamping blocks 12 move relative to each other and respectively engage with the two slots 10 of the chip frame 9, fixing the chip frame 9 in the positioning holes 13. In this embodiment, the positioning frame 11 is designed with three positioning holes 13, which are arranged equidistantly. In other embodiments not shown, the number of positioning holes 13 can be set to more or fewer, for example, one, two, four, five, etc., to simultaneously operate a corresponding number of sequencing chips 2. The number and position of the soaking reaction zones and the number and position of the slots 16 vary accordingly with the number of positioning holes 13.

[0105] In this embodiment, the grippers 7 of the transfer device 6 indirectly hold the sequencing chip 2 through the chip holding device to achieve the connection between the transfer device 6 and the sequencing chip 2. Indirectly holding the sequencing chip 2 can prevent cross-contamination when the grippers 7 hold different sequencing chips 2. In addition, the grippers 7 indirectly hold the sequencing chip 2 through the chip holding frame 9, so only the clamping structure needs to be processed on the chip frame 9, instead of processing the clamping structure on the sequencing chip 2, which reduces the processing cost of the sequencing chip 2 and maximizes the utilization of the surface area of ​​the sequencing chip 2.

[0106] Of course, in other embodiments not shown, the transfer device 6 and the sequencing chip 2 can also be connected by directly clamping the sequencing chip 2 with the gripper 7.

[0107] Preferably, the gene sequencing reaction apparatus 1 includes a protective cover, with the soaking container housed inside the cover. If environmental conditions permit, such as during aseptic operation, the sequencing reaction process can be completely open. However, in many cases, to avoid external interference, the sequencing reaction process needs to be carried out in a closed environment. Placing the soaking container inside the protective cover provides a sealed reaction environment, ensuring the quality of the gene sequencing reaction. The transfer device 6 can also be optionally placed inside the protective cover, depending on the requirements.

[0108] The air blowing device 25 is used to blow away chemical reagents from the surface of the sequencing chip 2 and / or the surface of the chip holder on which the sequencing chip is mounted. In this embodiment, the air blowing device 25 is used to blow away residual chemical reagents from the surfaces of the sequencing chip 2 and the chip frame 9. Blowing away residual chemical reagents from the surfaces of the sequencing chip 2 and the chip frame 9 can minimize cross-contamination when the sequencing chip 2 is immersed in the next immersion reaction zone.

[0109] See Figure 11 In this embodiment, the blowing device 25 includes a nozzle. The nozzle is used to spray gas, which blows away residual chemical reagents on the surface of the sequencing chip 2 and the chip frame 9.

[0110] The control device 23 is used to control and monitor the operation of the gene sequencing reaction device 1. In this embodiment, the control device 23 is coupled to the temperature control device 5 to control the temperature of the chemical reagents, and is also coupled to the transfer device 6 to control the soaking time and / or soaking sequence of the sequencing chip 2 in the soaking reaction zone.

[0111] In this embodiment, the control device 23 is a built-in control device, which is mounted on the support platform 8. In other embodiments not shown, an external control device may also be coupled to the gene sequencing reaction device 1 to control and monitor the operation of the gene sequencing reaction device 1.

[0112] By controlling the temperature control device 5 and the transfer device 6 through the control device 23, the gene sequencing reaction equipment 1 can be automated, thereby improving the quality and efficiency of gene sequencing.

[0113] like Figures 1 to 3 As shown, in this embodiment, the support platform 8 is a cabinet. The aforementioned soaking container, temperature control device 5, transfer device 6, upper material storage device 14, lower material storage device 15, container lid 4, flip-top mechanism 22, air blowing device 25, and control device 23 are all located on the top surface of the cabinet. The cabinet can store and hold reagents and tools required for gene sequencing reactions. Casters 27 are installed under the cabinet to facilitate movement of the gene sequencing reaction equipment 1.

[0114] In other embodiments not shown, the support platform 8 may also be a support plate.

[0115] like Figures 1 to 3 As shown, in this embodiment, an indicator light 28 can be added to the gene sequencing reaction device 1 to trigger an alarm when the gene sequencing reaction device 1 malfunctions.

[0116] In this embodiment, each soaking reaction zone contains a chemical reagent for gene sequencing reaction. The transfer device 6 holds a group of sequencing chips 2 and soaks them in a group of soaking reaction zones for a period of time, then transfers the group of sequencing chips 2 to the next group of soaking reaction zones for a period of time, and so on. Each sequencing chip 2 in this group of sequencing chips 2 can complete one sequencing reaction process after being soaked in multiple soaking reaction zones.

[0117] For example, in such Figures 1 to 3 In the illustrated embodiment, the sequencing chips are clamped in groups of three, numbered from right to left as chip 1, chip 2, chip 3, etc. Immersion tanks 3 are numbered sequentially from right to left, starting from the back row and continuing from left to right, as follows: Immersion tank 1 (rightmost in the back row), Immersion tank 2, Immersion tank 3, ..., Immersion tank 17 (leftmost in the back row), Immersion tank 18 (leftmost in the front row), Immersion tank 19, ..., Immersion tank 34 (rightmost in the front row). When the gene sequencing reaction begins, chip 3 is immersed in immersion tank 1. After immersion for a predetermined time, chip 3 is transferred to immersion tank 2 for further immersion, while chip 2 is immersed in immersion tank 1. After immersion for the predetermined time, chip 3 is transferred to immersion tank 3 for further immersion, while chip 2 is immersed in immersion tank 2, and chip 1 is immersed in immersion tank 1. Following this pattern, after a set of chips has been soaked in a set of soaking tanks, each sequencing chip 2 is sequentially transferred to a soaking tank with a number one higher than its own soaking tank to continue soaking, until all sequencing chips 2 have completed the corresponding soaking process in all soaking tanks. The soaking process in soaking tanks 16-17, 18-19, and 33-34 is similar to the soaking process in soaking tanks 1-2, allowing some sequencing chips 2 to be soaked while others are not.

[0118] In some embodiments, one or more soaking tanks at the head and / or tail of at least one row of soaking tanks (the specific number can be set according to the number of sequencing chips held each time) can be an empty container or contain the same chemical reagent as the adjacent soaking tank or contain a reagent that has no effect on the gene sequencing reaction, so that the sequencing chip 2 that does not participate in the soaking is also in the soaking tank.

[0119] In some embodiments, the soaking tanks (or soaking reaction zones) can be grouped, for example, according to the number of sequencing chips held at each time, and each group of soaking tanks (or soaking reaction zones) contains the same chemical reagent. Each time, a group of sequencing chips is soaked in a group of soaking tanks containing the same chemical reagent. This setup can improve the efficiency of gene sequencing reactions, but requires more soaking tanks (or soaking reaction zones).

[0120] The gene sequencing system of this embodiment includes a DNA sample loading device and a gene sequencing reaction device 1, wherein the gene sequencing reaction device 1 is the aforementioned gene sequencing reaction device 1.

[0121] The gene sequencing reaction method of this embodiment includes: adding chemical reagents for gene sequencing reaction into the soaking reaction zone of the soaking container; controlling the temperature of the chemical reagents in the soaking reaction zone; and immersing the sequencing chip 2, which has a DNA sample loading structure on its surface and is loaded with DNA sample, in the chemical reagents for a predetermined time and then taking it out.

[0122] Preferably, different chemical reagents for gene sequencing reactions are added to multiple soaking reaction zones, and the sequencing chip 2 is soaked in multiple soaking reaction zones in a predetermined order for a predetermined time.

[0123] The following combination Figures 1-3 Briefly describe the working process of gene sequencing reaction using the gene sequencing reaction device 1 and gene sequencing reaction method of the above embodiments.

[0124] 1. Initial state: Fill the soaking tank 3 with chemical reagents for gene sequencing reaction, adjust the heat-conducting liquid in the water bath to a suitable temperature, install the sequencing chip 2 to be subjected to gene sequencing reaction onto the chip frame 9, and place all chip frames 9 into the upper material storage device 14.

[0125] 2. The gripper 7 of the transfer device 6 holds the chip frame 9 and immerses each chip frame 9 in the soaking tank 3 for a period of time. When all the chip frames 9 have been soaked in a row of soaking tanks 3, the longitudinal moving mechanism 20 drives the transverse moving mechanism 19 to move longitudinally, so that each chip frame 9 can be soaked in another row of soaking tanks 3 until all the soaking tanks 3 have been soaked.

[0126] 3. After all the chip frames 9 have been soaked in the soaking tank 3, the transfer device 6 places the chip frames 9 into the lower part storage device 15. The DNA sample on the sequencing chip 2 completes the sequencing reaction process and is ready to be removed.

[0127] After the DNA sample on sequencing chip 2 completes the sequencing reaction process, it is transferred to an optical imaging device for imaging.

[0128] It should be understood that the above steps are only one possible working process that the gene sequencing reaction device 1 can perform, and do not mean that it can only perform these steps, nor are they intended to limit the scope of protection of this invention. Furthermore, since the chemical reactions involved in this invention are not within the scope of protection claimed by this invention, and not disclosing such content does not affect those skilled in the art's understanding of this invention, the above-mentioned chemical reactions are not disclosed herein.

[0129] The gene sequencing system and gene sequencing reaction method of the present invention have similar technical effects to the gene sequencing reaction device 1 of the present invention.

[0130] As can be seen from the above description, the above embodiments of the present invention can achieve at least one of the following technical effects:

[0131] Using this gene sequencing reaction device 1, gene sequencing reactions can be achieved by immersion.

[0132] Because the sequencing chip 2 is immersed in chemical reagents in different immersion reaction zones, it can complete all the steps required for the sequencing reaction.

[0133] The chemical reagents in the soaking reaction zone can be reused, thereby reducing the cost of consumables.

[0134] The immersion method eliminates the problem of uneven liquid flow rate, and bubbles are less likely to form on the surface of sequencing chip 2, ensuring a more uniform and complete chemical reaction.

[0135] The sequencing chip 2 is subjected to uniform liquid pressure and uniform heating in the immersion tank 3, so it will not deform.

[0136] It can soak multiple sequencing chips simultaneously, which has the advantage of high throughput.

[0137] It has no complex fluid system, fewer parts, and advantages such as easy assembly and low manufacturing cost.

[0138] The gene sequencing reaction equipment 1 is automatically controlled by the control device 23 to achieve automated operation.

[0139] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them; although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications can still be made to the specific implementation of the present invention or equivalent substitutions can be made to some technical features without departing from the spirit of the technical solutions of the present invention, and all such modifications and substitutions should be covered within the scope of the technical solutions claimed in the present invention.

Claims

1. A gene sequencing reaction device (1), characterized in that, include: An immersion container having an immersion reaction zone, the immersion reaction zone being used to hold chemical reagents for gene sequencing reactions and to immerse a sequencing chip (2) with a DNA sample loading structure on its surface and already loaded with DNA samples in the chemical reagents for gene sequencing reactions; and A transfer device is used to insert the sequencing chip (2) into or remove it from the immersion reaction zone; wherein The gene sequencing reaction apparatus (1) includes a plurality of said soaking containers; and / or, said soaking containers include a plurality of said soaking reaction zones isolated from each other; The transfer device (6) is configured to move the sequencing chip (2) in one of the soaking reaction zones into another soaking reaction zone.

2. The gene sequencing reaction apparatus (1) according to claim 1, characterized in that, It also includes a temperature control device for controlling the temperature of the chemical reagents in the soaking reaction zone.

3. The gene sequencing reaction apparatus (1) according to claim 1, characterized in that, The gene sequencing reaction device (1) also includes: Upper storage device (14) for placing the sequencing chip (2) to be used for gene sequencing reaction; and / or, The lower part storage device (15) is used to place the sequencing chip (2) that has completed the gene sequencing reaction.

4. The gene sequencing reaction apparatus (1) according to claim 1, characterized in that, The transfer device (6) includes a connector for connecting to the sequencing chip (2) and a motion mechanism that is driven to the connector to change the working position of the connector.

5. The gene sequencing reaction apparatus (1) according to claim 4, characterized in that, Multiple soaking reaction zones are arranged in one or more rows in the transverse direction; the motion mechanism includes a transverse moving mechanism (19), a longitudinal moving mechanism (20) and a vertical moving mechanism (21). The transverse moving mechanism (19) is located on the longitudinal moving mechanism (20), the vertical moving mechanism (21) is located on the transverse moving mechanism (19), and the connecting part is located on the vertical moving mechanism (21). The longitudinal moving mechanism (20) drives the transverse moving mechanism (19) to move longitudinally, the transverse moving mechanism (19) drives the vertical moving mechanism (21) to move laterally, and the vertical moving mechanism (21) drives the connecting part to move vertically.

6. The gene sequencing reaction apparatus (1) according to any one of claims 1 to 5, characterized in that, The gene sequencing reaction device (1) further includes a chip holding device, which includes one or more chip mounting positions (24), and the sequencing chip (2) is mounted on the chip mounting positions (24) to move the sequencing chip (2) by moving the chip holding device.

7. The gene sequencing reaction apparatus (1) according to any one of claims 1 to 5, characterized in that, The gene sequencing reaction device (1) also includes a control device (23), wherein, The gene sequencing reaction apparatus (1) further includes a temperature control device for controlling the temperature of the chemical reagents in the immersion reaction zone, and a control device (23) coupled to the temperature control device to control the temperature of the chemical reagents; and / or, The control device (23) is coupled to the transfer device (6) to control the soaking time and / or soaking sequence of the sequencing chip (2) in the soaking reaction zone.

8. The gene sequencing reaction apparatus (1) according to any one of claims 1 to 5, characterized in that, The gene sequencing reaction device (1) includes the sequencing chip (2), and the surface of the sequencing chip (2) has a DNA sample loading structure.

9. The gene sequencing reaction apparatus (1) according to any one of claims 1 to 5, characterized in that, The gene sequencing reaction device (1) includes a protective cover, and the immersion container is housed inside the protective cover.

10. The gene sequencing reaction apparatus (1) according to any one of claims 1 to 5, characterized in that, The gene sequencing reaction apparatus (1) includes a blowing device (25) for blowing away chemical reagents from the surface of the sequencing chip (2) and / or the surface of the chip holder on which the sequencing chip is mounted.

11. A gene sequencing system, comprising a DNA sample loading device and a gene sequencing reaction device (1), characterized in that, The gene sequencing reaction device (1) is the gene sequencing reaction device (1) according to any one of claims 1 to 10.

12. A gene sequencing reaction method using the gene sequencing reaction apparatus (1) according to any one of claims 1 to 10, characterized in that, The gene sequencing reaction method includes: Add chemical reagents for gene sequencing reaction to the soaking reaction zone of the soaking container; The sequencing chip (2) with a DNA sample loading structure on its surface and loaded with a DNA sample is immersed in the chemical reagent for a predetermined time and then removed. Different chemical reagents for gene sequencing reactions are added to multiple soaking reaction zones, and the sequencing chip (2) is soaked in the multiple soaking reaction zones for a predetermined time in a predetermined order.

13. The gene sequencing reaction method according to claim 12, characterized in that, The gene sequencing reaction method includes controlling the temperature of the chemical reagents within the immersion reaction zone.

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