Gene sequencing system and gene sequencing method

Abstract

Disclosed in the present invention is a gene sequencing system, comprising: a flow cell assembly, which is used for biochemical reactions; a first pipetting module, which is used for injecting samples and reagents into the flow cell assembly; a reuse module, which is used for liquid transfer; a second pipetting module, which is connected to the flow cell assembly and used for sucking up liquid from the flow cell assembly and injecting the liquid into the reuse module for transfer; and a waste liquid tank, which is used for waste liquid storage. The reuse module comprises a switching valve and a reuse liquid storage assembly. The reuse liquid storage assembly is provided with reuse liquid storage reservoirs corresponding to the reagents on a one-to-one basis. The switching valve is provided with a liquid inlet and several liquid outlets, the liquid inlet being connected to the second pipetting module, and the liquid outlets comprising a waste liquid port connected to the waste liquid tank and reuse ports corresponding to the reuse liquid storage reservoirs on a one-to-one basis, each reuse port being connected to a corresponding reuse liquid storage reservoir. Further disclosed in the present invention is a gene sequencing method.

Description

Gene sequencing system and gene sequencing method Technical Field

[0001] The present invention belongs to the field of in vitro diagnostic technology, and specifically relates to a gene sequencing system and a gene sequencing method. Background Art

[0002] Gene sequencing is a new genetic testing technology that can analyze and determine the complete sequence of genes from blood or saliva, predicting the likelihood of developing various diseases, individual behavioral characteristics, and the rationality of their behavior. Gene sequencing technology can identify individual disease genes, enabling early prevention and treatment.

[0003] In existing technologies, sequencers use nucleic acid samples placed on a solid-phase substrate for nucleic acid detection. However, each reagent is typically disposed of as waste after completing a single biochemical reaction. However, at this point, a high proportion of usable components are still present in the reagent, resulting in low actual reagent utilization. Disposing of reagents directly as waste after completing a single biochemical reaction is highly wasteful. Furthermore, because the reagents required for biochemical reactions are often expensive, low reagent utilization leads to high costs for gene sequencing, which undoubtedly increases user costs.

[0004] Summary of the Invention

[0005] In view of this, the purpose of the present invention is to provide a gene sequencing system and a gene sequencing method, based on the problem of low utilization rate of existing biochemical reaction reagents, to improve reagent utilization rate through reagent reuse, thereby reducing costs.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] The present invention first proposes a gene sequencing system, comprising:

[0008] a flow cell assembly for conducting biochemical reactions;

[0009] a first pipetting module, for injecting samples and reagents into the flow cell assembly;

[0010] a multiplexing module for transferring liquids;

[0011] a second pipetting module, connected to the flow cell assembly, and used to receive the liquid in the flow cell assembly and inject the liquid into the multiplexing module for transfer;

[0012] Waste liquid barrel, used to store waste liquid;

[0013] The reuse module includes a switching valve and a reuse liquid storage component, and the reuse liquid storage component is provided with a reuse liquid storage pool corresponding one to one with the reagent; the switching valve is provided with a liquid inlet and several liquid outlets, the liquid inlet is connected to the second pipetting module, and the liquid outlet includes a waste liquid port connected to the waste liquid barrel and a reuse port set one to one with the reuse liquid storage pool, and the reuse port is connected to the corresponding reuse liquid storage pool.

[0014] Furthermore, the waste liquid ports are arranged in one-to-one correspondence with the reuse ports, and the corresponding reuse ports and waste liquid ports are arranged adjacent to each other.

[0015] Furthermore, a hole blocking position is provided on the switching valve.

[0016] Furthermore, the liquid inlet of the switching valve is arranged at the center of the switching valve, and the liquid outlets are evenly distributed in a ring around the axis of the switching valve.

[0017] Furthermore, the flow cell assembly includes a flow cell, and reaction channels are arranged in parallel in the flow cell.

[0018] Furthermore, the flow cell assembly includes a liquid inlet manifold and a liquid outlet manifold respectively located at the liquid inlet end and the liquid outlet end of the flow cell.

[0019] Furthermore, the liquid inlet manifold includes a liquid inlet pipe and a liquid inlet branch pipe connected to the liquid inlet pipe, the liquid inlet branch pipes are arranged in a one-to-one correspondence with the reaction channels, and the liquid inlet branch pipes are connected to the corresponding reaction channels.

[0020] Furthermore, a valve is provided on the liquid inlet branch pipe.

[0021] Furthermore, the liquid outlet manifold includes a liquid outlet pipe and a liquid outlet branch pipe connected to the liquid outlet pipe, the liquid outlet branch pipes are arranged in a one-to-one correspondence with the reaction channels, and the liquid outlet branch pipes are connected to the corresponding reaction channels, and the liquid outlet pipe is connected to the second pipetting module.

[0022] Furthermore, the flow cell assembly also includes a reaction heating device for heating the flow cell.

[0023] Furthermore, the first pipetting module includes a reagent tank for storing reagents, a reagent needle assembly for aspirating or injecting reagents, and a pipetting drive assembly for driving the reagent needle assembly to move;

[0024] The reagent needle assembly includes a reagent needle and a first injection pump connected to the reagent needle. Under the action of the first injection pump, the reagent needle draws reagent from the reagent chamber and injects the reagent into the flow cell assembly.

[0025] Furthermore, the reagent needle includes a needle tube and a needle sleeve arranged outside the needle tube, the lower end of the needle tube extends out of the needle sleeve and forms a needle tip, and the lower end of the needle sleeve is provided with a tapered end.

[0026] Furthermore, a preheating device for preheating the reagent in the reagent needle is provided in the needle sleeve.

[0027] Furthermore, the first pipetting module also includes a liquid adding block, in which a liquid adding channel is provided; the liquid adding channel includes a sealing section for airtightly cooperating with the needle sleeve, a receiving section for accommodating the needle tip, and an output section for outputting the reagent, the sealing section is located above the output section, the receiving section is located between the output section and the sealing section, and a sealing ring is provided in the sealing section for sealingly cooperating with the tapered end of the needle sleeve.

[0028] Furthermore, a connector is provided at the lower end of the liquid adding block.

[0029] Furthermore, the first pipetting module also includes a first three-way valve, the inlet of which is connected to the liquid adding channel; one outlet of the first three-way valve is connected to the flow pool assembly, and the other outlet is connected to the waste liquid bucket.

[0030] Furthermore, the number of the reagent needle assembly is one or at least two.

[0031] Furthermore, the reagent needle assembly is set to at least two; the first pipetting module also includes a confluence assembly, the confluence assembly includes a docking tube arranged in a one-to-one correspondence with the reagent needle assembly, the upper end of the docking tube is connected to the liquid adding block, and the lower end of all the docking tubes is provided with a confluence plate, and the lowest position of the confluence plate is provided with a confluence outlet.

[0032] Furthermore, the first pipetting module also includes a first three-way valve, the inlet of the first three-way valve is connected to the confluence outlet; one outlet of the first three-way valve is connected to the flow pool assembly, and the other outlet is connected to the waste liquid bucket.

[0033] Furthermore, the pipetting drive assembly drives the reagent needle assembly to move along three directions perpendicular to each other, and the three directions include a vertical direction and two horizontal directions perpendicular to each other; or, the pipetting drive assembly drives the reagent needle assembly to move along a first horizontal direction and a vertical direction; and also includes a reagent tank drive assembly for driving the reagent tank to move along a second horizontal direction, and the first horizontal direction and the second horizontal direction are perpendicular to each other.

[0034] Furthermore, the reagent compartment is provided with a plurality of storage areas, the storage areas including a room temperature area for room temperature storage and a refrigerated area for refrigerated storage; or, the storage areas including a room temperature area for room temperature storage, a refrigerated area for refrigerated storage and a frozen area for frozen storage.

[0035] Furthermore, a turntable is provided in the storage area, and at least one annular liquid storage area is provided on the turntable, and each of the annular liquid storage areas is provided with at least one liquid storage level; or, at least one liquid storage level is provided at intervals along a straight line direction in the storage area; or, a plurality of liquid storage levels are provided in a rectangular array in the storage area.

[0036] Furthermore, a degassing module for degassing the reagent is provided between the reagent needle and the first injection pump.

[0037] Furthermore, the reuse liquid storage pool is located in the reagent compartment.

[0038] Furthermore, the first pipetting module further includes a cleaning assembly for cleaning the reagent needle; the cleaning assembly includes a cleaning bucket for storing cleaning liquid, an inner wall cleaning unit for cleaning the inner wall of the reagent needle, and an outer wall cleaning unit for cleaning the outer wall of the reagent needle;

[0039] The inner wall cleaning unit includes a second three-way valve and a first cleaning pump, wherein the two inlets of the second three-way valve are connected to the first injection pump and the first cleaning pump respectively, and one outlet of the second three-way valve is connected to the reagent needle; the liquid inlet of the first cleaning pump is connected to the cleaning bucket;

[0040] The outer wall cleaning unit includes a cleaning tank and a second cleaning pump. A cleaning liquid inlet is provided on the side wall of the cleaning tank. The liquid outlet of the second cleaning pump is connected to the cleaning liquid inlet, and the liquid inlet of the second cleaning pump is connected to the cleaning bucket.

[0041] Furthermore, a drain port is provided at the bottom of the cleaning pool, and a third cleaning pump is provided between the drain port of the cleaning pool and the waste liquid barrel.

[0042] Furthermore, the reagent needle includes a needle tube and a needle sleeve arranged outside the needle tube, the lower end of the needle tube extends out of the needle sleeve and forms a needle tip; a first air-drying device is provided above the cleaning pool, and a first air-drying hole for the reagent needle to pass through is provided in the first air-drying device, and the aperture of the first air-drying hole can be adjusted so that the first air-drying device has two states of "on" and "off"; when the first air-drying device is in the "on" state, the aperture of the first air-drying hole is the largest, and the first air-drying hole can be passed through by the needle sleeve; when the first air-drying device is in the "off" state, the aperture of the first air-drying hole is the smallest, and the first air-drying hole can allow the needle tip to pass through but serves as a barrier to the needle sleeve.

[0043] Furthermore, the first pipetting module also includes a liquid adding block, which is provided with a liquid adding channel; the reagent needle includes a needle tube and a needle sleeve arranged outside the needle tube, and the lower end of the needle tube extends out of the needle sleeve and forms a needle tip; the cleaning component includes a second air-drying device located above the liquid adding block, and the second air-drying device is provided with a second air-drying hole for the reagent needle to pass through, and the aperture of the second air-drying hole is adjustable so that the second air-drying device has two states of "on" and "off"; when the second air-drying device is in the "on" state, the aperture of the second air-drying hole is the largest, and the second air-drying hole can be passed through by the needle sleeve; when the second air-drying device is in the "off" state, the aperture of the second air-drying hole is the smallest, and the second air-drying hole can allow the needle tip to pass through but blocks the needle sleeve.

[0044] Furthermore, the cleaning component also includes a third air-drying device arranged on the horizontal moving path of the reagent needle, and the third air-drying device is provided with an air-drying channel located on the reagent needle path and through which the reagent needle can pass, and a third air pump for sucking or supplying air from the air-drying channel.

[0045] Furthermore, a check valve is provided at the liquid inlet of the first cleaning pump and the second cleaning pump respectively.

[0046] Furthermore, the second pipetting module includes a second syringe pump and a third three-way valve, and the third three-way valve enables the second syringe pump to communicate with the liquid inlet of the flow cell assembly or the switching valve.

[0047] Furthermore, the second injection pump is connected to the waste liquid barrel.

[0048] The present invention also proposes a gene sequencing method, comprising the following steps:

[0049] Step 1: Start gene sequencing, let i = 1;

[0050] Step 2: Determine whether the reused liquid reservoir corresponding to the i-th reagent stores the i-th reagent for reuse: if so, use the reagent needle to aspirate the i-th reagent in the reused liquid reservoir; if not, use the reagent needle to aspirate the i-th reagent in the corresponding storage position;

[0051] Step 3: Move the reagent needle to the liquid adding block and dock it with the liquid adding block; control the second syringe pump to connect to the flow cell through the third three-way valve;

[0052] Step 4: The first syringe pump drives the reagent needle to inject the i-th reagent, and the i-th reagent is injected into the flow cell through the liquid adding channel in the liquid adding block; at the same time, the original reagent in the flow cell is driven into the second syringe pump;

[0053] Step 5: The i-th reagent undergoes a biochemical reaction in the flow cell;

[0054] During this process, the third three-way valve is used to control the connection between the second syringe pump and the liquid inlet of the switching valve; it is determined whether the reagent in the second syringe pump meets the reuse conditions: if so, the switching valve is switched to the reuse port corresponding to the reagent, and the second syringe pump transfers the reagent to the corresponding reuse liquid storage tank through the switching valve; if not, the switching valve is switched to the waste liquid port, and the second syringe pump transfers the reagent to the waste liquid bucket through the switching valve;

[0055] During this process, it is determined whether the current gene sequencing task is completed: if so, the current sequencing task is completed; if not, i=i+1 is set and step 2 is executed in a loop.

[0056] Furthermore, in step 2, before the reagent needle absorbs the i-th reagent, the reagent needle needs to be cleaned, and the method steps for cleaning the reagent needle are:

[0057] 21) Move the reagent needle to the top of the cleaning tank and control the first air drying device to the "on" state;

[0058] 22) driving the reagent needle vertically downward into the cleaning tank;

[0059] 23) Turn on the second cleaning pump and use the outer wall cleaning unit to clean the outer wall of the reagent needle;

[0060] Switch the second three-way valve to connect the first cleaning pump to the reagent needle, start the first cleaning pump, and use the inner wall cleaning unit to clean the inner wall of the reagent needle;

[0061] 24) driving the reagent needle to move vertically upward so that the needle tip of the reagent needle is located within the first air-drying device;

[0062] 25) Controlling the first air drying device to be in the "off" state, turning on the first air pump to air dry the outer wall of the needle tip of the reagent needle, and turning off the first air pump after air drying;

[0063] 26) Drive the reagent needle to the corresponding multiplexing liquid reservoir or liquid storage level.

[0064] Furthermore, before the reagent needle draws the i-th reagent, the second three-way valve is switched to connect the first syringe pump to the reagent needle.

[0065] Furthermore, in step 26), when the reagent needle moves toward the corresponding reuse liquid storage pool or liquid storage level and passes through the air-drying channel, the third air pump is turned on to air-dry the outer wall of the reagent needle passing through the air-drying channel.

[0066] Furthermore, in step three, when the reagent needle moves toward the liquid adding block and passes through the air-drying channel, the third air pump is turned on to air-dry the outer wall of the reagent needle passing through the air-drying channel.

[0067] Furthermore, in step three, the process of docking the reagent needle with the liquid adding block is as follows: controlling the second air-drying device to be in the "off" state, driving the reagent needle to move downward and making the needle tip of the reagent needle located in the second air-drying device, and turning on the second air pump to air-dry the outer wall of the needle tip of the reagent needle; after the air-drying is completed, controlling the second air-drying device to be in the "on" state, and driving the reagent needle to continue to move downward to dock with the liquid adding block.

[0068] Furthermore, in step 4, the process of injecting the i-th reagent into the flow cell is as follows: switching the first three-way valve to connect with the waste liquid barrel, and the first injection pump driving the reagent needle to inject the i-th reagent; when the i-th reagent passes through the outlet of the first three-way valve connected to the waste liquid barrel, closing the first injection pump, and switching the first three-way valve to connect with the flow cell; the first injection pump drives the reagent needle to inject the i-th reagent, and after the i-th reagent enters the flow cell, closing the first injection pump.

[0069] Furthermore, after the reagent needle absorbs the i-th reagent, the i-th reagent is degassed at least once, and the process of each degassing treatment is: using the first injection pump to drive the i-th reagent in the reagent needle to move upward and pass through the degassing module, and then using the first injection pump to drive the i-th reagent in the reagent needle to move downward and pass through the degassing module again.

[0070] Furthermore, before the i-th reagent in the reagent needle is injected into the flow cell through the liquid adding block, the i-th reagent is heated to a preheating temperature lower than that required for the biochemical reaction.

[0071] Furthermore, during the biochemical reaction, the i-th reagent in the flow cell is heated to a desired reaction temperature.

[0072] Furthermore, in step 2, the process of using the reagent needle to draw the i-th reagent is:

[0073] (1) driving the reagent needle to draw a certain amount of push reagent through the first syringe pump;

[0074] (2) driving the reagent needle to absorb air through the first syringe pump, and forming a first air column in the reagent needle;

[0075] (3) driving the reagent needle to draw the i-th reagent by the first syringe pump;

[0076] (4) The reagent needle is driven by the first syringe pump to absorb air and form a second air column in the reagent needle.

[0077] Furthermore, the method further includes step 6: after the current sequencing detection task is completed, determining whether all sequencing detection tasks are completed: if so, executing step 7; if not, setting i=1, executing steps 2 and 9;

[0078] Step 7: Control the second syringe pump to communicate with the flow cell through the third three-way valve, and drive the reagent in the flow cell into the second syringe pump;

[0079] Step 8: Control the second syringe pump to connect to the liquid inlet of the switching valve through the third three-way valve to determine whether the reagent meets the reuse condition: if so, switch the switching valve to the reuse port corresponding to the reagent, and the second syringe pump transfers the reagent to the corresponding reuse liquid storage tank through the switching valve; if not, switch the switching valve to the waste liquid port, and the second syringe pump transfers the reagent to the waste liquid bucket through the switching valve; then execute step 9;

[0080] Step 9: Clean the piping system.

[0081] Furthermore, in step 1, when gene sequencing is started, the pipeline system is cleaned.

[0082] Furthermore, the method for cleaning the pipeline system is: first, use the first injection pump to drive the reagent needle to absorb the cleaning liquid from the cleaning pool; then, the reagent needle is docked with the liquid adding block, and the cleaning liquid in the reagent needle is injected into the second injection pump through the flow pool by the first injection pump; finally, use the second injection pump to discharge the cleaning liquid into the waste liquid barrel.

[0083] Furthermore, the method for cleaning the pipeline system is: first, connect the first cleaning pump to the reagent needle through the second three-way valve; and dock the reagent needle with the liquid adding block; control the second injection pump to be connected to the flow pool through the third three-way valve; then, start the first cleaning pump, and inject the cleaning liquid into the second injection pump through the reagent needle through the flow pool; finally, turn off the first cleaning pump, and use the second injection pump to discharge the cleaning liquid into the waste liquid barrel.

[0084] Furthermore, the reagent reuse condition is that the number of times the corresponding reagent participates in the biochemical reaction is less than or equal to a set number; or, the reagent reuse condition is that the nucleotide concentration in the corresponding reagent is not less than a set threshold.

[0085] Furthermore, in step 4, during the process of injecting the i-th reagent into the flow cell, the first syringe pump is started to apply a pushing force (positive pressure) to the i-th reagent, and the second syringe pump is started to apply a suction force (negative pressure) to the i-th reagent.

[0086] The beneficial effects of the present invention are:

[0087] The gene sequencing system of the present invention is provided with a first pipetting module, which uses the first pipetting module to inject the reagents required for the biochemical reaction into the flow cell assembly, and the biochemical reaction is carried out in the flow cell assembly; during the process of the first pipetting module injecting the reagents into the flow cell assembly, the original reagents in the flow cell assembly are transferred to the second pipetting module; during the process of the biochemical reaction in the flow cell assembly, the second pipetting module injects the reagents into the switching valve. Depending on the situation of the reagent, the switching valve is connected to the reuse port to inject the reagents into the reuse liquid storage pool for reuse, or the switching valve is connected to the waste liquid port to inject the reagents into the waste liquid bucket for waste liquid treatment. In this way, the gene sequencing system of the present invention can enable the reagents to participate in at least two biochemical reactions, which can not only effectively improve the utilization rate of the reagents and thus reduce costs, but also perform reagent reuse and transfer during the biochemical reaction process without affecting the sequencing efficiency.

[0088] The gene sequencing method of the present invention first determines whether the corresponding reuse liquid storage pool stores the i-th reagent to be reused before adding reagents. If so, the i-th reagent is drawn from the reuse liquid storage pool; if not, the i-th reagent is drawn from the storage position; then the reagent needle is docked with the liquid adding block, and the second injection pump is connected to the flow pool. The first injection pump is used to drive the i-th reagent to be injected into the flow pool, and the original reagent in the flow pool enters the second injection pump; finally, the i-th reagent participates in the biochemical reaction in the flow pool, and the second injection pump transfers the reagent therein to the corresponding reuse liquid storage pool or waste liquid barrel through the switching valve; in this way, reagent reuse can be achieved, and the reagent utilization rate is improved through reagent reuse, thereby reducing costs. BRIEF DESCRIPTION OF THE DRAWINGS

[0089] In order to make the purpose, technical solutions and beneficial effects of the present invention more clear, the present invention provides the following drawings for illustration:

[0090] FIG1 is a schematic diagram of an embodiment of a gene sequencing system of the present invention;

[0091] FIG2 is a schematic structural diagram of a flow cell assembly;

[0092] FIG3 is a schematic structural diagram of a first pipetting module;

[0093] Figure 4 is a schematic structural diagram of a reagent needle;

[0094] FIG5 is a schematic diagram of the coordination relationship between the reagent needle and the liquid adding block;

[0095] FIG6 is a schematic structural diagram of a first air-drying device;

[0096] FIG7 is a schematic structural diagram of a second air-drying device;

[0097] FIG8 is a schematic structural diagram of a third air-drying device;

[0098] FIG9 is a schematic structural diagram of a switching valve;

[0099] FIG10 is a flow chart of the gene sequencing method of the present invention.

[0100] Explanation of reference numerals: 100 - flow cell assembly; 101 - flow cell; 102 - reaction channel; 103 - liquid inlet manifold; 1031 - liquid inlet pipe; 1032 - liquid inlet branch pipe; 1033 - valve; 104 - liquid outlet manifold; 1041 - liquid outlet pipe; 1042 - liquid outlet branch pipe; 200 - first pipetting module; 201 - reagent needle; 2011 - needle tube; 2012 - needle cover; 2013 - needle tip; 2014 - tapered end; 202 - First injection pump; 203-liquid adding block; 2031-sealing ring; 2032-connector; 204-liquid adding channel; 2041-sealing section; 2042-accommodating section; 2043-output section; 205-first three-way valve; 206-butt joint; 207-manifold; 208-confluent outlet; 209-degassing module; 210-reagent compartment; 211-normal temperature zone; 212-refrigerated zone; 213-liquid storage level; 214-second three-way valve; 215-first cleaning pump; 2151-check valve; 216-cleaning tank; 21 61 - cleaning liquid inlet; 2162 - drain port; 217 - second cleaning pump; 2171 - check valve; 218 - third cleaning pump; 219 - first air-drying device; 2191 - first air-drying hole; 2192 - first air-drying module; 220 - first air pump; 221 - second air-drying device; 2211 - second air-drying hole; 2212 - second air-drying module; 222 - second air pump; 223 - third air-drying device; 2231 - air-drying channel; 224 - third air pump; 225 - push reagent tank; 226 - cleaning bucket; 300 - Second pipetting module; 301 - Second syringe pump; 302 - Third three-way valve; 400 - Reusing module; 401 - Switching valve; 402 - Reusing liquid reservoir; 403 - Liquid inlet; 404 - Waste liquid port; 405 - Reusing port; 406 - Plug hole; 500 - Waste liquid bucket. DETAILED DESCRIPTION

[0101] The present invention will be further described below with reference to the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it. However, the embodiments are not intended to limit the present invention.

[0102] As shown in Figure 1, the gene sequencing system of this embodiment includes a flow cell assembly 100, a first pipetting module 200, a second pipetting module 300, a multiplexing module 400 and a waste liquid bucket 500. Specifically, the flow cell assembly 100 is used to provide a place for performing biochemical reactions. The first pipetting module 200 is used to inject reagents into the flow cell assembly 100. The multiplexing module 400 is used to transfer liquids. The second pipetting module 300 is connected to the flow cell assembly 100, and is used to receive the liquid in the flow cell assembly 100 and inject the liquid into the multiplexing module 400 for transfer. The waste liquid bucket 500 is used to store waste liquid.

[0103] As shown in Figure 2, the flow cell assembly 100 of the present embodiment includes a flow cell 101, and a reaction channel 102 is arranged in parallel in the flow cell 101. Four reaction channels 102 are arranged in parallel in the flow cell 101 of the present embodiment. Specifically, the quantity of reaction channels 102 is provided according to actual needs, such as being set to 1, 2, 3, 5 and more than 5, and no further description is given. The flow cell assembly 100 of the present embodiment includes a liquid inlet manifold 103 and a liquid outlet manifold 104 positioned respectively at the liquid inlet end and the liquid outlet end of the flow cell 101. The liquid inlet manifold 103 of the present embodiment includes a liquid inlet pipe 1031 and a liquid inlet branch 1032 connected with the liquid inlet pipe 1031, and the liquid inlet branch 1032 is provided with a one-to-one correspondence with the reaction channel 102, and the liquid inlet branch 1032 is connected with the corresponding reaction channel 102. Like this, by a liquid inlet pipe 1031, reagent is injected into the corresponding reaction channel 102 respectively by the liquid inlet branch 1032. Specifically, the liquid inlet branch 1032 of this embodiment is provided with a valve 1033, which controls the opening and closing of the liquid inlet branch 1032 by the valve 1033, thereby controlling the reagent to selectively enter the corresponding reaction channel 102. The liquid outlet manifold 104 of this embodiment includes a liquid outlet pipe 1041 and a liquid outlet branch 1042 connected to the liquid outlet pipe 1041. The liquid outlet branch 1042 is arranged in a one-to-one correspondence with the reaction channel 102, and the liquid outlet branch 1042 is connected to the corresponding reaction channel 102, and the liquid outlet pipe 1041 is connected to the second pipetting module 300. In a preferred embodiment of this embodiment, the flow cell assembly 100 also includes a reaction heating device (not shown in the figure) for heating the flow cell 101. The reaction heating device is used to heat the reagents in the flow cell 101 to the temperature required for the biochemical reaction.

[0104] As shown in Figure 3, the first pipetting module 200 of this embodiment includes a reagent compartment 210 for storing reagents, a reagent needle assembly for drawing or injecting reagents, and a pipetting drive assembly for driving the reagent needle assembly to move. In this embodiment, the reagent needle assembly includes a reagent needle 201 and a first injection pump 202 connected to the reagent needle 201. Under the action of the first injection pump 202, the reagent needle 201 draws the reagent from the reagent compartment 210 and injects the reagent into the flow cell assembly 100. Specifically, as shown in Figure 4, the reagent needle 201 of this embodiment includes a needle tube 2011 and a needle sleeve 2012 that is sleeved outside the needle tube 2011. The lower end of the needle tube 2011 extends outside the needle sleeve 2012 and forms a needle tip 2013. The lower end of the needle sleeve 2012 is provided with a tapered end 2014. In a preferred embodiment of this embodiment, a preheating device (not shown) is provided within the needle sleeve 2012 for preheating the reagent within the reagent needle 2011. This preheating device allows the reagent within the reagent needle 2011 to be preheated to a temperature close to or equal to the biochemical reaction temperature. This allows the reagent to quickly reach the required biochemical reaction temperature within the flow cell 101 after being injected into the flow cell 101, thereby improving efficiency. In this embodiment, a push reagent reservoir 225 is connected to the first syringe pump 202, which contains the push reagent.

[0105] The first pipetting module 200 of this embodiment also includes a liquid adding block 203, which is provided with a liquid adding channel 204. The liquid adding channel 204 is connected to the flow cell 101. Specifically, the liquid adding channel is connected to the liquid inlet pipe 1031. The liquid adding block 203 is mainly used to dock with the reagent needle 201 so that the reagent needle 201 can inject the absorbed reagent into the flow cell 101 through the liquid adding block 203. As shown in Figure 5, in this embodiment, the liquid adding channel 204 includes a sealing section 2041 for airtightly cooperating with the needle guard 2012, a receiving section 2042 for accommodating the needle tip 2013, and an output section 2043 for outputting the reagent. The sealing section 2041 is located above the output section 2043, and the receiving section 2042 is located between the output section 2043 and the sealing section 2041. The sealing section 2041 is provided with a sealing ring 2031 for sealingly cooperating with the tapered end 2014 of the needle guard 2012. In this way, a sealed passage is formed between the needle tube 2011 and the reaction channel 102 during the process of the reagent needle 201 injecting the reagent into the flow cell 101. In this embodiment, a connector 2032 is provided at the lower end of the liquid adding block 204 to facilitate connection with the liquid inlet manifold 103 or other components connected to the liquid inlet manifold 103, thereby connecting the liquid adding channel 204 to the liquid inlet pipe 1031.

[0106] In this embodiment, the first pipetting module 200 further includes a first three-way valve 205, the inlet of which is connected to the liquid-adding channel 204. One outlet of the first three-way valve 205 is connected to the liquid inlet pipe 1031, and the other outlet is connected to the waste liquid bucket 500. Thus, by configuring the first three-way valve 205, liquid can be selectively directed to either the flow cell 101 or the waste liquid bucket 500.

[0107] Specifically, the reagent needle assembly is set to one or two. When the reagent needle assembly is set to at least two, the first pipetting module 200 also includes a confluence assembly, and the confluence assembly includes a butt joint pipe 206 that is arranged one by one with the reagent needle assembly, and the upper end of the butt joint pipe 206 connects the liquid adding block 203, and the lower ends of all butt joint pipes 206 are provided with a confluence plate 207, and the lowest position of the confluence plate 207 is provided with a confluence liquid outlet 208. In the present embodiment, the confluence liquid outlet 208 is connected with the first three-way valve 205. In the present embodiment, the reagent needle assembly is set to two, that is, in the present embodiment, the liquid adding flow channel 204 is connected with the first three-way valve 205 by the butt joint pipe 206, the confluence plate 207 and the confluence liquid outlet 208. Like this, reagent can be injected into the flow cell 101 respectively by a plurality of reagent needles 201, thereby can improve efficiency.

[0108] In this embodiment, in order to drive the reagent needle assembly to move so as to draw the reagent at the set position and inject the reagent into the liquid adding channel 204, the pipetting drive assembly of this embodiment drives the reagent needle assembly to move along three directions perpendicular to each other, and the three directions include a vertical direction and two horizontal directions perpendicular to each other. Of course, in some other embodiments, the pipetting drive assembly can also drive the reagent needle assembly to move along the first horizontal direction and the vertical direction, and at the same time, a reagent tank drive assembly is provided for driving the reagent tank 210 to move along the second horizontal direction. The first horizontal direction and the second horizontal direction are perpendicular to each other, and the technical purpose of driving the reagent needle 201 to move relative to the reagent tank 210 in three directions perpendicular to each other can also be achieved.

[0109] As shown in Figure 3, the reagent compartment 210 of this embodiment is provided with multiple storage areas. These storage areas include a room temperature area 211 for room temperature storage and a cold storage area 212 for refrigerated storage. Room temperature area 211 is used to store reagents that require room temperature storage, while cold storage area 212 is used to store reagents that require refrigerated storage. In other embodiments, the storage areas may include room temperature area 211 for room temperature storage, cold storage area 212 for refrigerated storage, and a freezing area for frozen storage, with the latter being used to store reagents that require refrigerated storage, such as lyophilized pellets. Liquid storage levels within the storage areas can be achieved in a variety of ways. In this embodiment, multiple liquid storage levels 213 are provided in a rectangular array within the storage area. In other embodiments, at least one liquid storage level 213 may be spaced along a straight line within the storage area. In still other embodiments, a turntable may be provided within the storage area, with at least one annular liquid storage area disposed thereon, each of which contains at least one liquid storage level.

[0110] In a preferred embodiment of this embodiment, a degassing module 209 for degassing the reagent is provided between the reagent needle 201 and the first syringe pump 202. The degassing process of the reagent using the degassing module 209 is as follows: after the first syringe pump 202 drives the reagent needle 201 to draw the reagent, the first syringe pump 202 drives the reagent in the reagent needle 201 through the degassing module 209, and then drives the reagent in the reagent needle 201 to flow back into the reagent needle 201 and through the degassing module 209. This process is repeated for at least one cycle through the degassing module 209 to remove bubbles in the reagent.

[0111] The first pipetting module 200 of the present embodiment also includes a cleaning assembly for cleaning reagent needle. As shown in Figure 3, the cleaning assembly includes a cleaning bucket 226 for storing cleaning solution, an inner wall cleaning unit for cleaning the inner walls of reagent needle 201 and an outer wall cleaning unit for cleaning the outer wall of reagent needle. Specifically, in the present embodiment, the inner wall cleaning unit includes a second three-way valve 214 and a first cleaning pump 215, two imports of the second three-way valve 214 are connected to the first injection pump 202 and the first cleaning pump 215 respectively, and an outlet of the second three-way valve 214 is connected to the reagent needle 201. The liquid inlet of the first cleaning pump 215 is connected to the cleaning bucket 226. Thus, when the inner wall of the reagent needle 201 needs to be cleaned, the first cleaning pump 215 is controlled to be connected to the reagent needle 201 through the second three-way valve 214, and the first injection pump 202 is disconnected from the reagent needle 201. The first cleaning pump 215 is used to draw cleaning fluid from the cleaning barrel 226 to clean the inner wall of the reagent needle 201. When the reagent needle 201 needs to transfer reagent, the first cleaning pump 215 is controlled to be disconnected from the reagent needle 201 through the second three-way valve 214, and the first injection pump 202 is connected to the reagent needle 201. The outer wall cleaning unit of this embodiment includes a cleaning tank 216 and a second cleaning pump 217. A cleaning fluid inlet 2161 is provided on the side wall of the cleaning tank 216. The liquid outlet of the second cleaning pump 217 is connected to the cleaning fluid inlet 2161, and the liquid inlet of the second cleaning pump 217 is connected to the cleaning barrel 226. When the outer wall of the reagent needle 201 needs to be cleaned, the needle tip 2013 of the reagent needle 201 is inserted into the cleaning tank 216. When the second cleaning pump 217 is used to pump cleaning liquid, a spring is formed in the cleaning tank 216, thereby cleaning the outer wall of the reagent needle 201. A drainage port 2162 is provided at the bottom of the cleaning tank 216. A third cleaning pump 218 is provided between the drainage port 2162 of the cleaning tank 216 and the waste liquid bucket 500. The third cleaning pump 218 can be used to quickly drain the liquid in the cleaning tank 216 into the waste liquid bucket 500. Of course, in this embodiment, when cleaning the inner wall of the reagent needle 201, it can also be done in the cleaning tank 216. That is, the cleaning liquid used to clean the inner wall of the reagent needle 201 can be discharged into the waste liquid bucket 500 through the drainage port 2162, so that the inner and outer walls of the reagent needle 201 are cleaned simultaneously. In this embodiment, a check valve 2151 is provided at the liquid inlet of the first cleaning pump 215, and a check valve 2171 is provided at the liquid inlet of the second cleaning pump 217 to prevent liquid from flowing back into the cleaning tank 216 and contaminating the cleaning liquid.

[0112] In a preferred embodiment of this embodiment, a first air-drying device 219 is provided above the cleaning tank 216. A first air-drying hole 2191 is provided within the first air-drying device 219 for passage of the reagent needle 201. The diameter of the first air-drying hole 2191 is adjustable, allowing the first air-drying device 219 to have two states: "open" and "closed." When the first air-drying device 219 is in the "open" state, the diameter of the first air-drying hole 2191 is at its largest, allowing the needle guard 2012 to pass through. When the first air-drying device 219 is in the "closed" state, the diameter of the first air-drying hole 2191 is at its smallest, allowing the needle tip 2013 to pass through while blocking the needle guard 2012. Thus, when cleaning the inner and outer walls of the reagent needle 201, the first air-drying device 219 is in the "open" state, allowing the reagent needle 201 to pass through the first air-drying hole 2191 and enter the cleaning tank 216. When the reagent needle 201 is removed from the cleaning tank 216, the outer wall of the reagent needle 201 can be dried by the first air-drying device 219. Specifically, when the reagent needle 201 moves upward and the needle tip 2013 is located in the first air-drying device 219, the first air-drying device 219 is controlled to be in the "off" state. At this time, the first air pump 220 is started to draw air from the first air-drying hole 2191 or inject air into the first air-drying hole 2191, and the air flow formed by the air is used to dry the needle tip 2013. Specifically, there are many ways to make the first air-drying device 219 have two states of "on" and "off". In this embodiment, the first air-drying device 219 includes two first air-drying modules 2192, the first air-drying hole 2191 is arranged between the two first air-drying modules 2192, and the two first air-drying modules 2192 move relative to each other to adjust the "on" and "off" states of the first air-drying device 219. When the first air-drying device 219 is in the "on" state, the distance between the two first air-drying modules 2192 is the largest, and the first air-drying hole 2191 allows the needle sleeve to pass through; when the first air-drying device 219 is in the "off" state, the distance between the two first air-drying modules 2192 is the smallest, and the first air-drying hole 2191 allows the needle tip 2013 to pass through but acts as a barrier to the needle sleeve 2012, as shown in Figure 6.

[0113] In a preferred embodiment of this embodiment, the cleaning assembly includes a second air-drying device 221 located above the liquid-adding block 203. A second air-drying hole 2211 is provided within the second air-drying device 221 for passage of the reagent needle. The diameter of the second air-drying hole 2211 is adjustable, allowing the second air-drying device 221 to have two states: "open" and "closed." When the second air-drying device 221 is in the "open" state, the diameter of the second air-drying hole 2211 is at its largest, allowing the needle guard 2012 to pass through. When the second air-drying device 221 is in the "closed" state, the diameter of the second air-drying hole 2211 is at its smallest, allowing the needle tip 2013 to pass through while blocking the needle guard 2012. During the docking process between the reagent needle 201 and the liquid adding block 203, the second air-drying device 221 can be first controlled to be in the "off" state, the needle tip 2013 can be placed in the second air-drying hole 2211, and the second air pump 222 can be activated to draw air from or inject air into the second air-drying hole 2211, so that the air flow formed by the air dries the needle tip 2013. After the outer wall of the needle tip 2013 is dried, the second air-drying device 221 can be controlled to be in the "on" state, so that the reagent needle 201 moves downward to dock with the liquid adding block 203. Similarly, in this embodiment, the second air-drying device 221 includes two second air-drying modules 2212, the second air-drying hole 2211 is arranged between the two second air-drying modules 2212, and the two second air-drying modules 2212 move relative to each other to adjust the "on" and "off" states of the second air-drying device 221; when the second air-drying device 221 is in the "on" state, the distance between the two second air-drying modules 2212 is the largest, and the second air-drying hole 2211 allows the needle sleeve 2012 to pass through; when the second air-drying device 221 is in the "off" state, the distance between the two second air-drying modules 2212 is the smallest, and the second air-drying hole 2211 allows the needle tip 2013 to pass through but serves as a blocking effect on the needle sleeve 2012, as shown in Figure 7.

[0114] In a preferred embodiment of this embodiment, the cleaning assembly further includes a third air-drying device 223 disposed on the horizontal moving path of the reagent needle 201. The third air-drying device 223 is provided with an air-drying channel 2231 located on the horizontal moving path of the reagent needle 201 and through which the reagent needle 201 can pass, and a third air pump 224 for sucking or delivering air from the air-drying channel 2231, as shown in Figure 8. Specifically, the reagent needle 201 can be controlled to have a fixed horizontal moving path when moving back and forth between the reagent chamber 210 and the liquid adding block 203. The third air-drying device 223 is disposed on this horizontal moving path. Each time the reagent needle 201 passes through the air-drying channel 2231, it is air-dried, further enhancing the air-drying effect. The air-drying process does not affect the movement of the reagent needle 201, thereby improving efficiency.

[0115] As shown in Figure 1, the second pipetting module 300 of this embodiment includes a second syringe pump 301 and a third three-way valve 302. The third three-way valve 302 connects the second syringe pump 301 with the liquid inlet of the flow cell assembly 100 or the switching valve 401. In this way, when the reagent needle 201 is docked with the liquid adding block 203 and the first reagent pump 102 drives the reagent needle 201 to inject the reagent into the flow cell 101, the third three-way valve 302 controls the second syringe pump 301 to connect with the flow cell assembly 100. In the process of the reagent in the reagent needle 201 flowing toward the flow cell 101, the reagent in the flow cell 101 flows toward the second syringe pump 301. When the reagent in the reagent needle 201 enters the flow cell 101, the original reagent in the flow cell 101 enters the second syringe pump 301. During the biochemical reaction of the reagents within the flow cell 101, the third three-way valve 302 controls the second syringe pump 301 to communicate with the switching valve, and the second syringe pump 301 is used to transfer the reagents within the flow cell 101 through the switching valve. Of course, when the first syringe pump 202 drives the reagents within the reagent needle 201 toward the flow cell 101, the first syringe pump 202 applies positive pressure to the reagents to drive the reagents within the reagent needle 201 toward the flow cell 101. In a preferred embodiment of this embodiment, the second syringe pump 301 can also apply negative pressure to the reagents to move the reagents toward the second syringe pump 301. This not only increases the reagent flow rate but also effectively reduces the liquid pressure within the flow cell 101. In this embodiment, the second syringe pump 301 is also connected to the waste liquid bucket 500, so that waste liquid can be directly discharged into the waste liquid bucket via the second syringe pump 301.

[0116] As shown in Figure 1, the reuse module 400 of this embodiment includes a switching valve 401 and a reuse liquid storage component. The reuse liquid storage component of this embodiment is provided with a reuse liquid storage tank 402 corresponding to each reagent; the switching valve 401 is provided with a liquid inlet 403 and a plurality of liquid outlets, and the liquid inlet 403 is connected to the second pipetting module 300, that is, the liquid inlet 403 is connected to the second injection pump 301 through the third three-way valve 302. In this embodiment, the liquid outlet includes a waste liquid port 404 connected to the waste liquid barrel 500 and a reuse port 405 corresponding to each reuse liquid storage tank 402, and the reuse port 405 is connected to the corresponding reuse liquid storage tank 402. In a preferred embodiment of this embodiment, the waste liquid port 404 and the reuse port 405 are provided in a one-to-one correspondence, and the corresponding reuse port 405 and the waste liquid port 404 are provided adjacent to each other, so that cross contamination between different reagents can be avoided. In a preferred embodiment of this embodiment, a hole blocking position 406 is further provided on the switching valve 401. When the switching valve 401 is switched to the hole blocking position 406, the liquid inlet 403 of the switching valve 401 is disconnected from the second injection pump 301. In this embodiment, the liquid inlet 403 of the switching valve 401 is set at the center of the switching valve 401, and the liquid outlets are evenly distributed in a ring around the axis of the switching valve 401. In a preferred implementation manner of this embodiment, the reuse liquid storage pool 402 is located in the reagent compartment 210, and there is no need to set up a separate storage position for the reuse liquid storage pool 402. As shown in Figure 9, in this embodiment, the switching valve 401 of this embodiment is provided with four reuse ports 405, four waste liquid ports 404 and one hole blocking position 406. Specifically, the reuse liquid storage pool 402 is also set to four accordingly.

[0117] The specific implementation of the gene sequencing method of the present invention is described in detail below in conjunction with the above-mentioned gene sequencing system.

[0118] As shown in FIG10 , the gene sequencing method of this embodiment includes the following steps:

[0119] Step 1: Start gene sequencing, let i=1.

[0120] Specifically, in this embodiment, when gene sequencing is started, the pipeline system is cleaned. There are many methods for cleaning the pipeline system, and this embodiment cites two methods. The first method for cleaning the pipeline is: first, use the first injection pump 202 to drive the reagent needle 201 to draw cleaning liquid from the cleaning pool 216; then, the reagent needle 201 is docked with the liquid adding block 203, and the second injection pump 301 is controlled to be connected to the flow pool 101 through the third three-way valve 302, and the cleaning liquid in the reagent needle 201 is injected into the second injection pump 301 through the flow pool 101 by the first injection pump 202; finally, the cleaning liquid is discharged into the waste liquid bucket 500 by the second injection pump 301. In the process of injecting the cleaning liquid in the reagent needle 201 into the flow pool 101 and the second injection pump 301, the first injection pump 202 applies a pushing force (positive pressure) to the cleaning liquid, and the second injection pump 301 applies a suction force (negative pressure) to the cleaning liquid. The second method for cleaning the pipeline is: first, connect the first cleaning pump 215 to the reagent needle 201 through the second three-way valve 214; and connect the reagent needle 201 to the liquid adding block 203; control the second injection pump 301 to be connected to the flow pool 101 through the third three-way valve 302; then, start the first cleaning pump 215, and inject the cleaning liquid into the second injection pump 301 through the reagent needle 201 and the flow pool 101; finally, turn off the first cleaning pump 215, and use the second injection pump 301 to discharge the cleaning liquid into the waste liquid barrel 500.

[0121] Step 2: Determine whether the reused i-th reagent is stored in the reuse liquid storage pool 402 corresponding to the i-th reagent: If yes, use the reagent needle 201 to aspirate the i-th reagent in the reuse liquid storage pool 402; if not, use the reagent needle 201 to aspirate the i-th reagent in the corresponding storage position 213.

[0122] Specifically, before the reagent needle 201 absorbs the i-th reagent, the reagent needle 201 needs to be cleaned, and the method steps for cleaning the reagent needle 201 are:

[0123] 21) Move the reagent needle 201 to the top of the cleaning tank 216 and control the first air-drying device 219 to be in the "on" state. At this time, the reagent needle 201 can pass through the first air-drying hole 2191.

[0124] 22) Drive the reagent needle 201 vertically downward into the cleaning tank 216.

[0125] 23) Turn on the second cleaning pump 217 to inject the cleaning liquid in the cleaning barrel 226 into the cleaning pool 216 through the cleaning liquid inlet 2161, forming a spring in the cleaning pool 216, and use the outer wall cleaning unit to clean the outer wall of the reagent needle 201.

[0126] The second three-way valve 214 is switched to connect the first cleaning pump 215 with the reagent needle 201, and the first cleaning pump 215 is started to inject the cleaning liquid in the cleaning barrel 226 into the reagent needle 201 through the second three-way valve 214 to clean the inner wall of the reagent needle 201, that is, the inner wall cleaning unit is used to clean the inner wall of the reagent needle 201.

[0127] 24) The reagent needle 201 is driven to move vertically upward so that the needle tip of the reagent needle 201 is located in the first air-drying device 219 .

[0128] 25) Control the first air-drying device 219 to be in the "off" state. At this time, only the needle tip 2013 can pass through the first air-drying hole 2191. Turn on the first air pump 220 to air-dry the outer wall of the needle tip 2013 of the reagent needle 201. After air-drying, turn off the first air pump 220.

[0129] 26) driving the reagent needle 201 to the corresponding reuse liquid reservoir 402 or liquid storage level 213. Specifically, in a preferred embodiment of this embodiment, while the reagent needle 201 is moving toward the corresponding reuse liquid reservoir 402 or liquid storage level 213 and passing through the air-drying channel 2231, the third air pump 224 is turned on to air-dry the outer wall of the reagent needle 201 passing through the air-drying channel 2231.

[0130] In addition, after the reagent needle 201 is cleaned, and before the reagent needle 201 absorbs the i-th reagent, the second three-way valve 214 is switched to connect the first syringe pump 202 to the reagent needle 201 .

[0131] In this embodiment, the process of using the reagent needle 201 to draw the i-th reagent is as follows:

[0132] (1) The first syringe pump 202 drives the reagent needle 201 to draw a certain amount of push reagent from the push reagent reservoir 225;

[0133] (2) The reagent needle 201 is driven by the first syringe pump 202 to quantitatively absorb air and form a first air column in the reagent needle 201;

[0134] (3) The reagent needle 201 is driven by the first syringe pump 202 to quantitatively absorb the i-th reagent;

[0135] (4) The first syringe pump 202 drives the reagent needle 201 to quantitatively absorb air, and a second air column is formed in the reagent needle 201.

[0136] In a preferred embodiment of this embodiment, after the reagent needle 201 draws the i-th reagent, the i-th reagent is degassed at least once. Each degassing process is as follows: the first syringe pump 202 drives the i-th reagent in the reagent needle 201 upward and through the degassing module 209, and then the first syringe pump 202 drives the i-th reagent in the reagent needle 201 downward and again through the degassing module 209. This process is repeated at least once to remove bubbles in the i-th reagent.

[0137] Step 3: Move the reagent needle 201 to the liquid adding block 203 and dock it with the liquid adding block 203; control the second syringe pump 301 to communicate with the flow cell 101 through the third three-way valve 302. Specifically, the process of docking the reagent needle 201 with the liquid adding block 203 is as follows: control the second air-drying device 221 to be in the "off" state, at which time only the needle tip 2013 can pass through the second air-drying hole 2211, drive the reagent needle 201 to move downward so that the needle tip 2013 of the reagent needle 201 is located in the second air-drying device 221, and start the second air pump 222 to air-dry the outer wall of the needle tip 2013 of the reagent needle 201; after the air-drying is completed, turn off the second air pump 222, control the second air-drying device 221 to be in the "on" state, at which time the entire reagent needle 201 can pass through the second air-drying hole 2211, and drive the reagent needle 201 to continue moving downward to dock with the liquid adding block 203.

[0138] Specifically, in a preferred implementation of this embodiment, when the reagent needle 201 moves toward the liquid adding block 203 and passes through the air drying channel 2231 , the third air pump 224 is turned on to air dry the outer wall of the reagent needle 201 passing through the air drying channel 2231 .

[0139] Step 4: The first syringe pump 202 drives the reagent needle 201 to inject the i-th reagent, and the i-th reagent is injected into the flow cell 101 through the liquid adding channel 204 in the liquid adding block 203 ; at the same time, the original reagent in the flow cell 101 is driven into the second syringe pump 301 .

[0140] Specifically, in this embodiment, the process of injecting the i-th reagent into the flow cell 101 is as follows: the first three-way valve 205 is switched to be connected to the waste liquid barrel 500, and the first syringe pump 202 drives the reagent needle 201 to inject the i-th reagent; when the i-th reagent passes through the outlet of the first three-way valve 205 connected to the waste liquid barrel 500, the first syringe pump 202 is closed, and the first three-way valve 205 is switched to be connected to the flow cell 101; the first syringe pump 202 is restarted to drive the reagent needle 201 to inject the i-th reagent, and after the i-th reagent enters the flow cell 101, the first syringe pump 202 is closed. In a preferred embodiment of this embodiment, during the process of injecting the i-th reagent into the flow cell 101, the first syringe pump 202 is activated to apply a pushing force (positive pressure) to the i-th reagent, and the second syringe pump 301 is activated to apply a suction force (negative pressure) to the i-th reagent. In this way, the flow rate of the i-th reagent can be increased and the pressure applied to the flow cell 101 can be reduced.

[0141] Specifically, in a preferred implementation of this embodiment, before the i-th reagent in the reagent needle 201 is injected into the flow cell 101 through the liquid adding block 203, the i-th reagent is heated to a preheating temperature lower than the preheating temperature required for the biochemical reaction. The preheating temperature can be slightly lower than the biochemical reaction temperature or can be equal to the biochemical reaction temperature. In this way, when the i-th reagent is injected into the flow cell 101, the i-th reagent can quickly reach the biochemical reaction temperature.

[0142] Step 5: The i-th reagent undergoes a biochemical reaction in the flow cell 101. During the biochemical reaction, the i-th reagent in the flow cell 101 is heated to a desired reaction temperature, so that the i-th reagent participates in the biochemical reaction under the set temperature environment.

[0143] Specifically, the process of the biochemical reaction of the i-th reagent in the flow cell 101 is as follows:

[0144] 11) Synthesis: The i-th nucleotide and the i-th polymerase perform a polymerization reaction to extend the sequencing primer, so that the i-th nucleotide is bound to the nucleic acid molecule;

[0145] 12) Taking photos: exciting the detectable label and collecting the signal of the detectable label;

[0146] 13) Cleavage: The blocking group and detectable label of the i-th nucleotide are removed using a cleavage reagent.

[0147] During this process, the third three-way valve 302 controls the second syringe pump 301 to be connected to the liquid inlet of the switching valve 401; determines whether the reagent in the second syringe pump meets the reuse condition: if so, the switching valve is switched to the reuse port corresponding to the reagent, and the second syringe pump transfers the reagent to the corresponding reuse liquid storage tank through the switching valve; if not, the switching valve is switched to the waste liquid port, and the second syringe pump transfers the reagent to the waste liquid bucket through the switching valve;

[0148] During this process, it is determined whether the current gene sequencing task is completed: if so, the current sequencing detection task is completed; if not, set i=i+1 and loop through step 2.

[0149] Step 6: After the current sequencing detection task is completed, determine whether all sequencing detection tasks are completed: if so, execute step 7; if not, set i=1 and execute steps 2 and 9.

[0150] Step 7: Control the second syringe pump 301 to communicate with the flow cell 101 through the third three-way valve 302 , and drive the reagent in the flow cell 101 to enter the second syringe pump 301 .

[0151] Step 8: Control the second injection pump 301 to be connected to the liquid inlet 403 of the switching valve 401 through the third three-way valve 302, and determine whether the reagent meets the reuse conditions: if so, switch the switching valve 401 to the reuse port 405 corresponding to the reagent, and the second injection pump 301 transfers the reagent to the corresponding reuse liquid storage tank 402 through the switching valve 401; if not, switch the switching valve 401 to the waste liquid port 404, and the second injection pump 301 transfers the reagent to the waste liquid barrel 500 through the switching valve 401; execute step 9.

[0152] Specifically, the reagent reuse condition may be that the number of times the corresponding reagent participates in the biochemical reaction is less than or equal to a set number; of course, the reagent reuse condition may also be that the nucleotide concentration in the corresponding reagent is not less than a set threshold.

[0153] Step 9: Clean the piping system. The method for cleaning the piping system is the same as step 1 and will not be repeated here.

[0154] In this embodiment, the i-th reagent includes the i-th nucleotide and the i-th polymerase. Each gene sequencing task requires two to four biochemical reactions, with the nucleotides and polymerases involved in each biochemical reaction being different. In this embodiment, the biochemical reaction process and number of reactions in each gene sequencing task are comparable to those in the prior art and are not further described.

[0155] The above embodiments are merely preferred embodiments for the purpose of fully illustrating the present invention, and the scope of protection of the present invention is not limited thereto. Equivalent substitutions or modifications made by those skilled in the art based on the present invention are within the scope of protection of the present invention. The scope of protection of the present invention shall be subject to the claims.

Claims

1. A gene sequencing system, characterized in that: include: a flow cell assembly for conducting biochemical reactions; A first pipetting module, used for injecting samples and reagents into the flow cell assembly; a multiplexing module for transferring liquids; A second liquid transfer module is connected to the flow cell assembly and is used to receive the liquid in the flow cell assembly and inject the liquid into the multiplexing module for transfer; Waste liquid barrel, used to store waste liquid; The reuse module includes a switching valve and a reuse liquid storage component, and the reuse liquid storage component is provided with a reuse liquid storage tank corresponding to the reagent one by one; the switching valve is provided with a liquid inlet and a plurality of liquid outlets, the liquid inlet is connected to the second pipetting module, and the liquid outlet includes a waste liquid port connected to the waste liquid barrel and a reuse port arranged one by one with the reuse liquid storage tank, and the reuse port is connected to the corresponding reuse liquid storage tank.

2. The gene sequencing system according to claim 1, characterized in that: The waste liquid ports are arranged in one-to-one correspondence with the reuse ports, and the corresponding reuse ports and waste liquid ports are arranged adjacent to each other.

3. The gene sequencing system according to claim 1, characterized in that: The switching valve is also provided with a hole blocking position.

4. The gene sequencing system according to claim 1, characterized in that: The liquid inlet of the switching valve is arranged at the center of the switching valve, and the liquid outlets are evenly distributed in a ring around the axis of the switching valve.

5. The gene sequencing system according to claim 1, characterized in that: The flow cell assembly comprises a flow cell, and reaction channels are arranged in parallel in the flow cell.

6. The gene sequencing system according to claim 5, characterized in that: The flow cell assembly includes an inlet manifold and an outlet manifold respectively located at the inlet end and the outlet end of the flow cell.

7. The gene sequencing system according to claim 6, characterized in that: The liquid inlet manifold includes a liquid inlet pipe and a liquid inlet branch pipe connected to the liquid inlet pipe. The liquid inlet branch pipes are arranged in a one-to-one correspondence with the reaction channels, and the liquid inlet branch pipes are connected to the corresponding reaction channels.

8. The gene sequencing system according to claim 7, characterized in that: The liquid inlet branch pipe is provided with a valve.

9. The gene sequencing system according to claim 6, characterized in that: The liquid outlet manifold includes a liquid outlet pipe and a liquid outlet branch pipe connected to the liquid outlet pipe, the liquid outlet branch pipes are arranged in a one-to-one correspondence with the reaction channels, and the liquid outlet branch pipes are connected to the corresponding reaction channels, and the liquid outlet pipe is connected to the second pipetting module.

10. The gene sequencing system according to claim 5, characterized in that: The flow cell assembly further comprises a reaction heating device for heating the flow cell.

11. The gene sequencing system according to claim 1, characterized in that: The first pipetting module includes a reagent tank for storing reagents, a reagent needle assembly for aspirating or injecting reagents, and a pipetting drive assembly for driving the reagent needle assembly to move; The reagent needle assembly comprises a reagent needle and a first injection pump connected to the reagent needle. Under the action of the first injection pump, the reagent needle draws reagent from the reagent chamber and injects the reagent into the flow cell assembly.

12. The gene sequencing system according to claim 11, characterized in that: The reagent needle comprises a needle tube and a needle sleeve arranged outside the needle tube, the lower end of the needle tube extends out of the needle sleeve to form a needle tip, and the lower end of the needle sleeve is provided with a tapered closing end.

13. The gene sequencing system according to claim 12, characterized in that: A preheating device for preheating the reagent in the reagent needle is arranged in the needle sleeve.

14. The gene sequencing system according to claim 12, characterized in that: The first pipetting module also includes a liquid adding block, in which a liquid adding channel is provided; the liquid adding channel includes a sealing section for airtightly cooperating with the needle sleeve, a receiving section for accommodating the needle tip, and an output section for outputting the reagent, the sealing section is located above the output section, the receiving section is located between the output section and the sealing section, and a sealing ring for sealingly cooperating with the tapered end of the needle sleeve is provided in the sealing section.

15. The gene sequencing system according to claim 14, characterized in that: The lower end of the liquid adding block is provided with a connector.

16. The gene sequencing system according to claim 14, characterized in that: The first pipetting module also includes a first three-way valve, an inlet of which is connected to the liquid adding channel; one outlet of the first three-way valve is connected to the flow pool assembly, and the other outlet is connected to the waste liquid bucket.

17. The gene sequencing system according to claim 11, characterized in that: The reagent needle assembly is provided as one or at least two.

18. The gene sequencing system according to claim 14, characterized in that: The number of the reagent needle assemblies is at least two; the first pipetting module further comprises a confluence assembly, the confluence assembly comprising docking tubes arranged one-to-one with the reagent needle assemblies, the upper ends of the docking tubes being connected to the liquid adding blocks, the lower ends of all the docking tubes being provided with a confluence plate, and a confluence outlet being provided at the lowest position of the confluence plate.

19. The gene sequencing system according to claim 18, characterized in that: The first pipetting module also includes a first three-way valve, an inlet of the first three-way valve is connected to the confluence outlet; one outlet of the first three-way valve is connected to the flow pool assembly, and the other outlet is connected to the waste liquid bucket.

20. The gene sequencing system according to claim 17 or 18, characterized in that: The pipetting drive assembly drives the reagent needle assembly to move along three directions that are perpendicular to each other, and the three directions include a vertical direction and two horizontal directions that are perpendicular to each other; or, the pipetting drive assembly drives the reagent needle assembly to move along a first horizontal direction and a vertical direction; and also includes a reagent chamber drive assembly for driving the reagent chamber to move along a second horizontal direction, and the first horizontal direction and the second horizontal direction are perpendicular to each other.

21. The gene sequencing system according to claim 11, characterized in that: The reagent compartment is provided with a plurality of storage areas, the storage areas including a normal temperature area for normal temperature storage and a cold storage area for cold storage; or, the storage areas including a normal temperature area for normal temperature storage, a cold storage area for cold storage and a freezing area for frozen storage.

22. The gene sequencing system according to claim 21, characterized in that: A turntable is provided in the storage area, and at least one annular liquid storage area is provided on the turntable, and each of the annular liquid storage areas is provided with at least one liquid storage level; or, at least one liquid storage level is provided at intervals along a straight line direction in the storage area; or, a plurality of liquid storage levels are provided in a rectangular array in the storage area.

23. The gene sequencing system according to claim 11, characterized in that: A degassing module for degassing the reagent is provided between the reagent needle and the first injection pump.

24. The gene sequencing system according to claim 11, characterized in that: The multiplexing liquid storage pool is located in the reagent compartment.

25. The gene sequencing system according to claim 11, characterized in that: The first liquid transfer module further comprises a cleaning component for cleaning the reagent needle; the cleaning component comprises a cleaning bucket for storing cleaning liquid, an inner wall cleaning unit for cleaning the inner wall of the reagent needle, and an outer wall cleaning unit for cleaning the outer wall of the reagent needle; The inner wall cleaning unit comprises a second three-way valve and a first cleaning pump, wherein two inlets of the second three-way valve are respectively connected to the first injection pump and the first cleaning pump, and one outlet of the second three-way valve is connected to the reagent needle; and a liquid inlet of the first cleaning pump is connected to the cleaning barrel; The outer wall cleaning unit comprises a cleaning tank and a second cleaning pump. A cleaning liquid inlet is provided on the side wall of the cleaning tank. The liquid outlet of the second cleaning pump is connected to the cleaning liquid inlet. The liquid inlet of the second cleaning pump is connected to the cleaning bucket.

26. The gene sequencing system according to claim 25, characterized in that: A liquid discharge port is provided at the bottom of the cleaning pool, and a third cleaning pump is provided between the liquid discharge port of the cleaning pool and the waste liquid barrel.

27. The gene sequencing system according to claim 25, characterized in that: The reagent needle includes a needle tube and a needle sleeve arranged outside the needle tube, the lower end of the needle tube extends out of the needle sleeve and forms a needle tip; a first air-drying device is provided above the cleaning pool, and a first air-drying hole for the reagent needle to pass through is provided in the first air-drying device, and the aperture of the first air-drying hole can be adjusted so that the first air-drying device has two states of "on" and "off"; when the first air-drying device is in the "on" state, the aperture of the first air-drying hole is the largest, and the first air-drying hole can be passed through by the needle sleeve; when the first air-drying device is in the "off" state, the aperture of the first air-drying hole is the smallest, and the first air-drying hole can allow the needle tip to pass through but blocks the needle sleeve.

28. The gene sequencing system according to claim 25, characterized in that: The first pipetting module also includes a liquid adding block, in which a liquid adding channel is provided; the reagent needle includes a needle tube and a needle sleeve mounted outside the needle tube, the lower end of the needle tube extends out of the needle sleeve and forms a needle tip; the cleaning component includes a second air-drying device located above the liquid adding block, in which a second air-drying hole for the reagent needle to pass is provided, the aperture of the second air-drying hole is adjustable so that the second air-drying device has two states of "on" and "off"; when the second air-drying device is in the "on" state, the aperture of the second air-drying hole is the largest, and the second air-drying hole can be passed through by the needle sleeve; when the second air-drying device is in the "off" state, the aperture of the second air-drying hole is the smallest, and the second air-drying hole can be passed through by the needle tip but blocks the needle sleeve.

29. The gene sequencing system according to claim 25, characterized in that: The cleaning component also includes a third air-drying device arranged on the horizontal moving path of the reagent needle, and the third air-drying device is provided with an air-drying channel located on the reagent needle path and through which the reagent needle can pass, and a third air pump for sucking or delivering air from the air-drying channel.

30. The gene sequencing system according to claim 25, characterized in that: The first cleaning pump and the second cleaning pump are respectively provided with a check valve at the liquid inlet.

31. The gene sequencing system according to claim 1, characterized in that: The second pipetting module includes a second injection pump and a third three-way valve, and the third three-way valve enables the second injection pump to communicate with the liquid inlet of the flow cell assembly or the switching valve.

32. The gene sequencing system according to claim 31, characterized in that: The second injection pump is connected to the waste liquid barrel.

33. A gene sequencing method, characterized in that: The steps include: Step 1: Start gene sequencing, let i = 1; Step 2: Determine whether the reused liquid storage pool corresponding to the i-th reagent stores the i-th reagent for reuse: if so, use the reagent needle to absorb the i-th reagent in the reused liquid storage pool; if not, use the reagent needle to absorb the i-th reagent in the corresponding storage position; Step 3: Move the reagent needle to the liquid adding block and dock it with the liquid adding block; control the second injection pump to communicate with the flow cell through the third three-way valve; Step 4: The first injection pump drives the reagent needle to inject the i-th reagent, and the i-th reagent is injected into the flow cell through the liquid adding channel in the liquid adding block; at the same time, the original reagent in the flow cell is driven to enter the second injection pump; Step 5: the i-th reagent undergoes a biochemical reaction in the flow cell; In this process, the third three-way valve is used to control the second injection pump to be connected to the liquid inlet of the switching valve; it is determined whether the reagent in the second injection pump meets the reuse condition: if so, the switching valve is switched to the reuse port corresponding to the reagent, and the second injection pump transfers the reagent to the corresponding reuse liquid storage tank through the switching valve; if not, the switching valve is switched to the waste liquid port, and the second injection pump Transfer the reagents to the waste tank via the switching valve; In this process, it is determined whether the current gene sequencing task is completed: if so, the current sequencing detection task is completed; if not, i=i+1 is set and step 2 is executed in a loop.

34. The gene sequencing method according to claim 33, characterized in that: In step 2, before the reagent needle absorbs the i-th reagent, the reagent needle needs to be cleaned, and the method steps for cleaning the reagent needle are: 21) Move the reagent needle to the top of the cleaning tank and control the first air drying device to be in the "on" state; 22) driving the reagent needle to extend into the cleaning tank in a vertical downward direction; 23) Turn on the second cleaning pump and use the outer wall cleaning unit to clean the outer wall of the reagent needle; The second three-way valve is switched to connect the first cleaning pump to the reagent needle, and the first cleaning pump is started to clean the inner wall of the reagent needle using the inner wall cleaning unit; 24) driving the reagent needle to move in a vertically upward direction so that the needle tip of the reagent needle is located in the first air-drying device; 25) Control the first air-drying device to be in the "off" state, start the first air pump to air-dry the outer wall of the needle tip of the reagent needle, and turn off the first air pump after air-drying; 26) Drive the reagent needle to the corresponding multiplexing liquid storage tank or storage level.

35. The gene sequencing method according to claim 34, characterized in that: Before the reagent needle draws the i-th reagent, the second three-way valve is switched to connect the first syringe pump to the reagent needle.

36. The gene sequencing method according to claim 33 or 34, characterized in that: In the step 26), when the reagent needle moves toward the corresponding reuse liquid storage pool or liquid storage position and passes through the air-drying channel, the third air pump is turned on to air-dry the outer wall of the reagent needle passing through the air-drying channel.

37. The gene sequencing method according to claim 33, characterized in that: In the step three, when the reagent needle moves toward the liquid adding block and passes through the air drying channel, the third air pump is turned on to air dry the outer wall of the reagent needle passing through the air drying channel.

38. The gene sequencing method according to claim 33 or 37, characterized in that: In step three, the process of docking the reagent needle with the liquid adding block is as follows: control the second air-drying device to be in the "off" state, drive the reagent needle to move downward and make the needle tip of the reagent needle located in the second air-drying device, and start the second air pump to air-dry the outer wall of the needle tip of the reagent needle; after air-drying is completed, control the second air-drying device to be in the "on" state, and drive the reagent needle to continue to move downward to dock with the liquid adding block.

39. The gene sequencing method according to claim 33, characterized in that: In the step 4, the process of injecting the i-th reagent into the flow pool is as follows: the first three-way valve is switched to be connected to the waste liquid barrel, and the first injection pump drives the reagent needle to inject the i-th reagent; when the i-th reagent passes through the outlet of the first three-way valve connected to the waste liquid barrel, the first injection pump is closed, and the first three-way valve is switched to be connected to the flow pool; the first injection pump drives the reagent needle to inject the i-th reagent, and after the i-th reagent enters the flow pool, the first injection pump is closed.

40. The gene sequencing method according to claim 33, characterized in that: After the reagent needle absorbs the i-th reagent, the i-th reagent is degassed at least once, and the process of each degassing treatment is: using the first injection pump to drive the i-th reagent in the reagent needle to move upward and pass through the degassing module, and then using the first injection pump to drive the i-th reagent in the reagent needle to move downward and pass through the degassing module again.

41. The gene sequencing method according to claim 33, characterized in that: Before the i-th reagent in the reagent needle is injected into the flow cell through the liquid adding block, the i-th reagent is heated to a preheating temperature lower than that required for the biochemical reaction.

42. The gene sequencing method according to claim 33 or 41, characterized in that: During the biochemical reaction, the i-th reagent in the flow cell is heated to a desired reaction temperature.

43. The gene sequencing method according to claim 33, characterized in that: In step 2, the process of using the reagent needle to draw the i-th reagent is: (1) driving the reagent needle to absorb a certain amount of push reagent through the first syringe pump; (2) driving the reagent needle to absorb air through the first syringe pump, and forming a first air column in the reagent needle; (3) driving the reagent needle to draw the i-th reagent through the first syringe pump; (4) The reagent needle is driven by the first injection pump to absorb air and form a second air column in the reagent needle.

44. The gene sequencing method according to claim 33, characterized in that: The step also includes step 6: after the current sequencing detection task is completed, determine whether all sequencing detection tasks are completed: if so, execute step 7; if not, set i=1, and execute steps 2 and 9; Step 7: Control the second syringe pump to communicate with the flow cell through the third three-way valve, and drive the reagent in the flow cell to enter the second syringe pump; Step 8: Control the second syringe pump to communicate with the liquid inlet of the switching valve through the third three-way valve to determine whether the reagent meets the reuse condition: if so, switch the switching valve to the reuse port corresponding to the reagent, and the second syringe pump transfers the reagent to the corresponding reuse liquid storage tank through the switching valve; if not, switch the switching valve to the waste liquid port, and the second syringe pump transfers the reagent to the waste liquid barrel through the switching valve; execute step 9; Step 9: Clean the piping system.

45. The gene sequencing method according to claim 33, characterized in that: In the step 1, when the gene sequencing is started, the pipeline system is cleaned.

46. ​​The gene sequencing method according to claim 44 or 45, characterized in that: The method for cleaning the pipeline system is: first, use the first injection pump to drive the reagent needle to absorb the cleaning liquid from the cleaning pool; then, dock the reagent needle with the liquid adding block, and use the first injection pump to inject the cleaning liquid in the reagent needle into the second injection pump through the flow pool; finally, use the second injection pump to discharge the cleaning liquid into the waste liquid barrel.

47. The gene sequencing method according to claim 44 or 45, characterized in that: The method for cleaning the pipeline system is: first, connect the first cleaning pump with the reagent needle through the second three-way valve; and connect the reagent needle with the liquid adding block; control the second injection pump to be connected with the flow pool through the third three-way valve; then, start the first cleaning pump, and inject the cleaning liquid into the second injection pump through the reagent needle through the flow pool; finally, close the first cleaning pump, and use the second injection pump to discharge the cleaning liquid into the waste liquid barrel.

48. The gene sequencing method according to claim 33, characterized in that: The reagent reuse condition is that the number of times the corresponding reagent participates in the biochemical reaction is less than or equal to the set number of times; or, the reagent reuse condition is that the nucleotide concentration in the corresponding reagent is not less than the set threshold.

49. The gene sequencing method according to claim 33, characterized in that: In the step 4, during the process of injecting the i-th reagent into the flow cell, the first injection pump is started to apply a pushing force to the i-th reagent, and the second injection pump is started to apply a suction force to the i-th reagent.

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