Automatic liquid replenishing device and method for sequencer

Abstract

An automatic liquid replenishing device and method for a sequencer. In the automatic liquid replenishing device, a first connector is arranged above a sequencer and is communicated with a liquid inlet of a sequencing chip; a second connector is communicated with a first liquid storage device; a third connector is communicated with a second liquid storage device; a fourth connector is communicated with a third liquid storage device; two ends of a first connecting pipe are respectively communicated with the first connector and the second connector; two ends of a second connecting pipe are respectively communicated with the first connector and the third connector; two ends of a third connecting pipe are respectively communicated with the third connector and the fourth connector; a fourth connecting pipe is communicated with the fourth connector; a first peristaltic pump is arranged on the path of the first connecting pipe; and a second peristaltic pump is arranged on the path of the fourth connecting pipe.

Description

Automatic liquid replenishing device and method for sequencer Technical Field

[0001] The present application relates to the field of gene sequencing technology, for example, to an automatic liquid replenishing device and method for a sequencer. Background Art

[0002] Gene sequencing is a new genetic testing technology currently being used for microbial species identification, whole genome sequencing, and new species discovery. It can analyze and determine gene sequences from human appendages like blood and saliva, predicting the likelihood of developing various diseases, such as cancer or leukemia. A gene sequencer is an instrument used for gene sequencing. Currently, most gene sequencers use sequencing chips as their testing carriers. Gene molecules / nucleic acid molecules are injected onto the sequencing chip. Reagents flow across the functional surface of the sequencing chip, chemically reacting with the gene molecules and emitting specific signals. The gene sequencer achieves this detection by detecting light signals, electrical signals, and pH changes.

[0003] Summary of the Invention

[0004] The present application provides an automatic liquid replenishment device and method for a sequencer.

[0005] The present application discloses an automatic liquid replenishing device for a sequencer, comprising:

[0006] The liquid reservoir comprises at least a first liquid reservoir, a second liquid reservoir and a third liquid reservoir;

[0007] Connectors, comprising at least a first connector, a second connector, a third connector, and a fourth connector, wherein the first connector is disposed above the sequencer and communicates with the liquid inlet of the sequencing chip, the second connector is disposed at the opening of the first liquid reservoir and communicates with the first liquid reservoir, the third connector is disposed at the opening of the second liquid reservoir and communicates with the second liquid reservoir, and the fourth connector is disposed at the opening of the third liquid reservoir and communicates with the third liquid reservoir;

[0008] connecting tubes, comprising at least a first connecting tube, a second connecting tube, a third connecting tube, and a fourth connecting tube, wherein both ends of the first connecting tube are respectively in communication with the first connector and the second connector, both ends of the second connecting tube are respectively in communication with the first connector and the third connector, both ends of the third connecting tube are respectively in communication with the third connector and the fourth connector, and the fourth connecting tube is in communication with the fourth connector;

[0009] The peristaltic pump comprises at least a first peristaltic pump and a second peristaltic pump, wherein the first peristaltic pump is arranged on the path of the first connecting tube, and the second peristaltic pump is arranged on the path of the fourth connecting tube.

[0010] The present application discloses an automatic liquid replenishment method for a sequencer, which is applied to the automatic liquid replenishment device for the sequencer. The method comprises:

[0011] When replenishing a single solution, the first peristaltic pump is started to alternately squeeze the first connecting tube. As the gas in the first connecting tube is emptied, the liquid in the first liquid reservoir passes through the second connector and the first connecting tube to the first connector, and the liquid drips into the liquid inlet of the sequencing chip in the form of droplets. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] FIG1 is a schematic diagram of an automatic refill device for a sequencer provided in an embodiment of the present application;

[0013] FIG2 is a second schematic diagram of an automatic refill device for a sequencer provided in an embodiment of the present application;

[0014] FIG3 is a third schematic diagram of an automatic refill device for a sequencer provided in an embodiment of the present application;

[0015] FIG4 is a fourth schematic diagram of an automatic refill device for a sequencer provided in an embodiment of the present application;

[0016] FIG5 is a fifth schematic diagram of an automatic refill device for a sequencer provided in an embodiment of the present application;

[0017] FIG6 is a schematic diagram of a first connector provided in an embodiment of the present application;

[0018] FIG7 is a schematic diagram of a second connector provided in an embodiment of the present application;

[0019] FIG8 is a schematic diagram of a third connector provided in an embodiment of the present application;

[0020] FIG9 is a schematic diagram of a fourth connector provided in an embodiment of the present application;

[0021] FIG10 is a sequencing signal of a sequencer provided in an embodiment of the present application without rehydration;

[0022] FIG11 is a sequencing signal during the rehydration process of the sequencer provided in an embodiment of the present application;

[0023] FIG12 is a schematic structural diagram of a first connector provided in an embodiment of the present application;

[0024] FIG13 is a schematic structural diagram of a second connector provided in an embodiment of the present application;

[0025] FIG14 is a cross-sectional view of the second connector in FIG13;

[0026] FIG15 is a schematic structural diagram of another second connector provided in an embodiment of the present application;

[0027] FIG16 is a cross-sectional view of another second connector provided in an embodiment of the present application;

[0028] FIG17 is a schematic structural diagram of another second connector provided in an embodiment of the present application;

[0029] FIG18 is a cross-sectional view of a second connector with a hollow structure provided by an embodiment of the present application;

[0030] FIG19 is a cross-sectional view of another second connector with a hollow structure provided by an embodiment of the present application;

[0031] FIG20 is a cross-sectional view of another second connector with a hollow structure provided in an embodiment of the present application.

[0032] In the figure: 11. First liquid reservoir; 12. Second liquid reservoir; 13. Third liquid reservoir; 21. First connector; 22. Second connector; 23. Third connector; 24. Fourth connector; 201. Connecting body; 2011. Mounting hole; 2012. Exhaust hole; 2013. Guide surface; 2014. Conical groove; 2015. Cylindrical groove; 202. Capillary; 203. Extension tube; 204. Limiting portion; 205. Sealing portion; 31. First connecting tube; 32. Second connecting tube; 33. Third connecting tube; 34. Fourth connecting tube; 41. First peristaltic pump; 42. Second peristaltic pump; 50. Incubator; 100. Sequencer; 101. Sequencing chip. DETAILED DESCRIPTION

[0033] The following describes in detail embodiments of the present application. Examples of the embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present application, and should not be construed as limiting the present application.

[0034] In the description of this application, unless otherwise expressly specified or limited, the terms "connected," "connected," and "fixed" should be understood in a broad sense. For example, they can refer to fixed connections, detachable connections, or integration; mechanical connections or electrical connections; direct connections or indirect connections through an intermediate medium; and can refer to internal communication between two components or interaction between two components. Those skilled in the art will understand the specific meanings of the above terms in this application based on the specific circumstances.

[0035] In this application, unless otherwise expressly specified or limited, a first feature being "above" or "below" a second feature may include the first and second features being in direct contact, or may include the first and second features being in contact not directly but through another feature between them. Moreover, a first feature being "above," "above," and "above" a second feature may include the first feature being directly above or obliquely above the second feature, or may simply mean that the first feature is higher in level than the second feature. A first feature being "below," "below," and "below" a second feature may include the first feature being directly below or obliquely below the second feature, or may simply mean that the first feature is lower in level than the second feature.

[0036] The technical solution of the present application will be further explained below with reference to the accompanying drawings and through specific implementation methods.

[0037] During sequencing, long sequencing times are accompanied by reagent consumption, library replenishment, and buffer replacement, requiring sample addition, fluid replenishment, and fluid replacement on the detection device. Most of these operations use manual fluid replenishment methods, which wastes labor costs, has low work efficiency, a high error rate, and is prone to missed replenishment.

[0038] Although there are electric mechanical valve and solenoid valve rehydration systems to replace manual rehydration, the system needs to be equipped with a vacuum pump or a pressure pump, and the opening and closing of the liquid valve is controlled by an automatic control system to drive the liquid inflow. The structure is complex and the volume is large, the operation is cumbersome, the work efficiency is low, the failure rate is high, and liquid is easily left at the valve, causing pollution and difficulty in cleaning.

[0039] In single-molecule sequencing technology, because each molecule needs to be detected one by one, the consistency of the sequencing biochemical system is required to be high. However, the reagent capacity provided in the sequencing chip is limited. Long-term sequencing will be accompanied by reagent consumption and pH changes, and it is difficult to keep the biochemical system consistent for a long time. Therefore, it is necessary to replenish the library and replace the buffer to restore the sequencing system and maintain the sequencing system in a relatively stable stage. Replenishing the library and changing the liquid will involve adding samples, replenishing the liquid, and changing the liquid in the detection device. Most of these operations use manual methods to replenish the liquid, resulting in a waste of labor costs and reduced work efficiency, and there are cases of missed replenishment and wrong replenishment.

[0040] Referring to Figures 1 to 5, an embodiment of the present application provides an automatic liquid replenishment device for a sequencer, which is mainly used for third-generation single-molecule long-read length sequencers, and can also be used for other types of sequencers. As long as the sequencer requires the actions of liquid injection, liquid replenishment, and liquid replacement, the automatic liquid replenishment device provided by this embodiment can be used.

[0041] The automatic fluid replenishment device includes at least one liquid reservoir, at least one connector, at least one connecting tube and at least one peristaltic pump. The liquid reservoir, connector, connecting tube and peristaltic pump are connected in a coordinated manner. The connector is used to connect the liquid reservoir and the connecting tube. The peristaltic pump is used to provide power for liquid flow. It can automatically replenish a single solution and can also automatically replenish an instant mixed solution. It can automatically replenish and stop replenishing liquid at any time.

[0042] For example, at least one liquid reservoir includes at least a first liquid reservoir 11, a second liquid reservoir 12, and a third liquid reservoir 13, and at least one connector includes at least a first connector 21, a second connector 22, a third connector 23, and a fourth connector 24. The first connector 21 is arranged above the sequencer 100 and is connected to the liquid inlet of the sequencing chip 101. The second connector 22 is arranged at the opening of the first liquid reservoir 11 and is connected to the first liquid reservoir 11. The third connector 23 is arranged at the opening of the second liquid reservoir 12 and is connected to the second liquid reservoir 12. The fourth connector 24 is arranged at the opening of the third liquid reservoir 13 and is connected to the third liquid reservoir 13. At least one connector The connecting pipe includes at least a first connecting pipe 31, a second connecting pipe 32, a third connecting pipe 33 and a fourth connecting pipe 34, the two ends of the first connecting pipe 31 are respectively connected to the first connector 21 and the second connector 22, the two ends of the second connecting pipe 32 are respectively connected to the first connector 21 and the third connector 23, the two ends of the third connecting pipe 33 are respectively connected to the third connector 23 and the fourth connector 24, and the fourth connecting pipe 34 is connected to the fourth connector 24; at least one peristaltic pump includes at least a first peristaltic pump 41 and a second peristaltic pump 42, the first peristaltic pump 41 is arranged on the path of the first connecting pipe 31, and the second peristaltic pump 42 is arranged on the path of the fourth connecting pipe 34.

[0043] In one embodiment, referring to FIG. 2 , when a single solution is replenished, the first peristaltic pump 41 is started to alternately squeeze the first connecting tube 31 . As the gas in the first connecting tube 31 is evacuated, the liquid in the first liquid reservoir 11 passes through the second connector 22 and the first connecting tube 31 to the first connector 21 , and the liquid drips into the liquid inlet of the sequencing chip 101 in the form of droplets.

[0044] In one embodiment, referring to Figures 3-5, during instant mixed solution replenishment, the second peristaltic pump 42 rotates forward, pumping liquid from the second reservoir 12 through the third connector 23, the third connecting tube 33, and the fourth connector 24 into the third reservoir 13. After the liquid in the second reservoir 12 is completely pumped out, the second peristaltic pump 42 rotates reversely, pushing the mixed solution in the third reservoir 13 through the fourth connector 24, the third connecting tube 33, the second reservoir 12, the third connector 23, and the second connecting tube 32 to the first connector 21. The liquid then drips into the inlet of the sequencing chip 101 in a droplet-like manner. This droplet-like manner ensures a relatively uniform and trace amount of liquid into the inlet of the sequencing chip 101, preventing the sequencing signal from being affected by excessive fluid flow or severe vibration.

[0045] In one embodiment, a peristaltic pump is a fluid transport device that pumps fluid by alternately squeezing and releasing the pump head on a connecting tube. For example, a six-roller peristaltic pump, including a stepper motor, can be used. This peristaltic pump allows for programmable pumping speed, volume, and time, ensuring uniform and accurate fluid delivery. This allows for automated, timed, and quantitative fluid delivery, effectively avoiding the drawbacks of manual fluid delivery, reducing labor waste, and lowering costs.

[0046] In one embodiment, the connecting tube is a commonly used silicone hose. The liquid reservoir can be a commonly used microcentrifuge tube (Eppendorf tube) or a threaded tube. All liquid reservoirs are arranged in an insulated box 50, and a low-temperature environment suitable for storing liquid reagents is formed in the insulated box 50. The supplementary liquid can be a sequencing buffer, a library, and other reagents related to sequencing. In this embodiment, the first liquid reservoir 11 stores sequencing buffer, which can be replenished separately; the second liquid reservoir 12 stores sequencing buffer, and the third liquid reservoir 13 stores the library. The library and sequencing buffer cannot be mixed in advance and placed for a long time, and need to be mixed immediately when added.

[0047] In one embodiment, referring to Figures 6 to 9 , the connector includes a connector body 201 with at least one mounting hole 2011 defined therein. A capillary tube 202 extends through the mounting hole 2011. Both ends of the capillary tube 202 protrude from the connector body 201, connecting the corresponding liquid reservoir and the connecting tube. Furthermore, the capillary tube 202 of the first connector 21 communicates with the liquid inlet of the sequencing chip 101, enabling liquid dripping.

[0048] In one embodiment, the number of mounting holes 2011 provided on each connector body 201 is determined by their location and function. In this embodiment, the first connector 21 is provided with two mounting holes 2011, with a capillary tube 202 provided for each mounting hole 2011. One capillary tube 202 communicates with the first connecting tube 31, and the other capillary tube 202 communicates with the second connecting tube 32. The second connector 22 is provided with one mounting hole 2011, with a capillary tube 202 provided for each mounting hole 2011. The capillary tube 202 communicates with the first connecting tube 31. The third connector 23 is provided with two mounting holes 2011, with a capillary tube 202 provided for each mounting hole 2011. One capillary tube 202 communicates with the second connecting tube 32, and the other capillary tube 202 communicates with the third connecting tube 33. Two mounting holes 2011 are provided on the fourth connector 24 . A capillary tube 202 is provided corresponding to each mounting hole 2011 . One capillary tube 202 is communicated with the third connecting tube 33 , and the other capillary tube 202 is communicated with the fourth connecting tube 34 .

[0049] In some embodiments, the connection body 201 may be in the shape of a cone to prevent the connector from completely entering the reservoir, thereby facilitating plugging and unplugging.

[0050] Some connectors have vent holes 2012 on their connecting body 201. These vent holes 2012 communicate with the outside atmosphere, allowing for exhaust and pressure relief, thereby balancing the internal and external pressures of the connector. The diameter of the vent holes 2012 can be adjusted to meet practical needs, for example, to 1.0 mm. In this embodiment, the vent holes 2012 are provided on the first connector 21 to ensure that the pressure in the cavity formed by the liquid inlet of the sequencing chip 101 and the first connector 21 is equal to the external atmospheric pressure. This allows liquid flowing out of the capillary 202 of the first connector 21 to drip into the liquid inlet of the test chip as droplets, where it is then driven into the reaction chamber of the test chip by the difference in gravitational potential energy.

[0051] The second connector 22 is provided with an exhaust hole 2012 to ensure that the pressure of the cavity of the first liquid reservoir 11 is equal to the external atmospheric pressure, so as to prevent the formation of negative pressure in the first liquid reservoir 11 which is not conducive to the extraction of liquid.

[0052] In this embodiment, the capillary tube 202 is a stainless steel tube. The capillary tube and the connecting body 201 are tightly sealed due to the frosted sidewalls of the mounting hole 2011 and the slight elasticity of the material, eliminating the need for screws or other auxiliary components. The connecting body 201 can be made of materials such as polyoxymethylene (POM), polymethyl dimethacrylate (PMMA), silicone resins such as polydimethylsiloxane (PDMS), polystyrene (PS) resins, polyethylene resins (such as polyvinyl chloride (PVC) or isobutyl polyethylene (TPS)), phenolic resins, epoxy resins, unsaturated polyester resins, polyurethanes, base-catalyzed polycaprolactam, or any copolymers thereof, such as copolymers of methyl methacrylate and polystyrene (MAS) and copolymers of methyl methacrylate and styrene (SAN).

[0053] In one embodiment, some connectors further include an extension tube 203, which is disposed within the liquid reservoir and communicates with the capillary tube 202. The extension tube 203 extends the capillary tube 202 so that it extends into the liquid in the liquid reservoir, for example, to the bottom of the liquid reservoir, allowing for more liquid to be drawn out and preventing corrosion of the capillary tube 202. The extension tube 203 is detachable for easy regular replacement. In this embodiment, the capillary tube 202 on the second connector 22 is provided with an extension tube 203, the two capillary tubes 202 on the third connector 23 are both provided with an extension tube 203, and the two capillary tubes 202 on the fourth connector 24 are provided with an extension tube 203. The capillary tube 202 connected to the third connecting tube 33 is provided with an extension tube 203, while the capillary tube 202 connected to the fourth connecting tube 34 is not provided with an extension tube 203. Since the fourth connecting tube 34 is used for gas flow, an extension tube 203 is not required to prevent liquid from entering the fourth connecting tube 34.

[0054] In this embodiment, the extension tube 203 is a silicone tube to prevent corrosion. The extension tube 203 can also be a plastic tube or a tube made of other corrosion-resistant materials.

[0055] When connecting the connecting tube to the capillary tube 202, the connecting tube is tightly fitted around the outside of the capillary tube 202 using its own elasticity, thereby achieving a sealed connection between the connecting tube and the capillary tube 202. When connecting the extension tube 203 to the capillary tube 202, the extension tube is tightly fitted around the outside of the capillary tube 202 using its own elasticity, thereby achieving a sealed connection between the extension tube 203 and the capillary tube 202.

[0056] 1 to 9 , an embodiment of the present application further provides an automatic refill method for a sequencer.

[0057] When replenishing a single solution, the first peristaltic pump 41 is started to alternately squeeze the first connecting tube 31. As the gas in the first connecting tube 31 is evacuated, the liquid in the first liquid reservoir 11 passes through the second connector 22 and the first connecting tube 31 to the first connector 21, and the liquid drips into the liquid inlet of the sequencing chip 101 in the form of drops.

[0058] When replenishing the mixed solution immediately, the second peristaltic pump 42 rotates forward to draw the liquid in the second liquid reservoir 12 through the third connector 23, the third connecting tube 33 and the fourth connector 24 into the third liquid reservoir 13. After the liquid in the second liquid reservoir 12 is completely drawn out, the second peristaltic pump 42 reverses to push the mixed solution in the third liquid reservoir 13 through the fourth connector 24, the third connecting tube 33, the second liquid reservoir 12, the third connector 23 and the second connecting tube 32 to reach the first connector 21. The liquid drips into the liquid inlet of the sequencing chip 101 in the form of droplets.

[0059] Before replenishing a single solution, preparations are made. For example, the speed, preset start time, and preset refill volume of the first peristaltic pump 41 are set. Sequencing buffer is then added to the first reservoir 11 in advance, ensuring that the first connecting tube 31 is free of liquid. When the preset start time is reached, the first peristaltic pump 41 automatically starts and, by alternately squeezing the first connecting tube 31, expel the air in the first connecting tube 31 through the capillary 202 on the first connector 21. As the air is expelled, the liquid in the first reservoir 11 flows through the capillary 202 on the second connector 22 into the first connecting tube 31, passes through the pump head of the first peristaltic pump 41, and flows along the first connecting tube 31 into the sequencing chip 101. As shown by the arrows in FIG2 , once all the air in the first connecting tube 31 has been expelled, the liquid begins to drip into the sequencing chip 101. When the liquid injection reaches the preset refill volume, the first peristaltic pump 41 automatically stops, thus achieving the purpose of automatic refill.

[0060] The first connecting tube 31 is divided into an inlet tube and an outlet tube by the first peristaltic pump 41. The inlet tube is located on one side of the first peristaltic pump 41 and connected to the second connector 22, and the outlet tube is located on the other side of the first peristaltic pump 41 and connected to the first connector 21. The longer the total length of the inlet tube and the outlet tube, the longer the emptying time, that is, the longer it takes to discharge the gas in the tube. Therefore, the length of the first connecting tube 31 can be determined according to the actual situation of the sequencer 100.

[0061] In this embodiment, the lengths of the inlet and outlet tubes are 42 mm and 62 mm, respectively, with an inner diameter of 0.5 mm and an outer diameter of 3 mm. The speed of the peristaltic pump's roller can be adjusted; the faster the speed, the faster the liquid inflow. When the speed is 50 rpm, if the preset refill volume is 400 μL, the emptying time and refill time are 62 ± 1 s and 45 ± 1 s, respectively, for a total time of 107 ± 3 s. If the preset refill volume is 1000 μL, the emptying time and refill time are 62 ± 1 s and 111 ± 2 s, respectively, for a total time of 173 ± 4 s. The refill accuracy for both 400 μL and 1000 μL volumes is less than 1%.

[0062] After a long period of sequencing, it is necessary to add two liquids, such as a library and a sequencing buffer, etc. However, before adding, in order to avoid the two solutions from mixing in advance and causing biochemical reactions, they need to be placed separately before adding. In this embodiment, the sequencing buffer with a larger sample volume is first loaded into the second liquid reservoir 12, and the library with a smaller sample volume is loaded into the third liquid reservoir 13. The speed, preset start time and preset rehydration volume of the second peristaltic pump 42 are set in advance. When the time reaches the preset start time, the second peristaltic pump 42 automatically starts to draw the liquid in the second liquid reservoir 12 into the third liquid reservoir 13. As shown by the arrow in Figure 3, since the extension tube 203 is provided on the third connector 23, it can ensure that the liquid in the second liquid reservoir 12 is completely extracted into the third liquid reservoir 13, so that mixing is sufficient. Since the capillary 202 connected to the fourth connecting tube 34 on the fourth connector 24 is not provided with an extension tube 203, it can avoid touching the liquid and prevent the liquid from being drawn out of the third liquid reservoir 13.

[0063] As shown in FIG4 , the liquid in the second liquid reservoir 12 is completely drawn into the third liquid reservoir 13 . As shown in FIG5 , the reverse function of the second peristaltic pump 42 is utilized to push the mixed liquid in the third liquid reservoir 13 from the third liquid reservoir 13 into the sequencing chip 101 , passing through the fourth connector 24 , the third connecting tube 33 , the third connector 23 , and the second connecting tube 32 , as indicated by the arrows in FIG5 .

[0064] By the above method, the timed quantitative replenishment and replacement of one or more liquids can be satisfied. In actual use, the number of liquid reservoirs can be increased as needed, and connectors, connecting tubes and peristaltic pumps can be increased accordingly.

[0065] The entire rehydration or liquid replacement process does not affect the normal operation of the sequencer 100, that is, the rehydration or liquid replacement has no effect on the sequencing current signal. Referring to Figures 10 and 11, by intercepting the sequencing signal of the sequencer 100 before rehydration and the sequencing signal during the rehydration process for comparison, it can be found that there is no obvious difference in the sequencing state in the two stages, no noise increases, no instability and other phenomena occur. Therefore, the rehydration process has almost no effect on the sequencing signal, proving that the rehydration or liquid replacement process does not interfere with the sequencing signal. Figures 10 and 11 in this embodiment only show the special signal of the single-molecule sequencer, which is an electrical signal; other mainstream sequencers mainly use optical signal detection methods. Although the signal cannot be extended to other types of sequencers, no matter whether the sequencer uses optical, electrical or chemical principles, as long as the actions of liquid addition, rehydration and liquid replacement are required, the automatic rehydration device provided by this embodiment can be used.

[0066] The dimensions of each connector and the dimensions of the capillary 202 on the connector can be set according to actual needs. The geometric dimensions of the first connector 21 match the liquid inlet of the sequencing chip 101. The second connector 22, the third connector 23, and the fourth connector 24 respectively match the openings of the corresponding liquid reservoirs. In this embodiment, for the first connector 21, the outer diameter of the capillary 202 is 0.65 mm, the outer diameter of the first connector 21 is 17.19 mm, and the inner diameter is 13.6 mm. For the second connector 22, the third connector 23, and the fourth connector 24, the outer diameter of the capillary 202 is 0.7 mm, and the length of the connecting body 201 is 14 mm.

[0067] In one embodiment, as shown in Figure 12 , first connector 21 is cylindrical for ease of processing and production. Because first connector 21 delivers liquid to sequencing chip 101, and the liquid drips into the liquid inlet of sequencing chip 101 in the form of droplets, first connector 21 can be configured as a hollow structure to provide space for the liquid to drip.

[0068] Among the above-mentioned connectors, since the second connector 22, the third connector 23 and the fourth connector 24 are all connected to the liquid reservoir, except for the number of mounting holes 2011 and exhaust holes 2012, the structures of the second connector 22, the third connector 23 and the fourth connector 24 are basically the same.

[0069] The following description will be made by taking the second connector 22 as an example, and the structures of other connectors may also refer to the second connector 22 .

[0070] For example, referring to Figures 1 and 13 to 17 , a limit portion 204 is provided at one end of the connecting body 201 near the connecting tube. When the connecting body 201 is inserted into the opening of the liquid reservoir, the limit portion 204 is located outside the opening and abuts the liquid reservoir. The provision of the limit portion 204 prevents the connecting body 201 from completely entering the liquid reservoir, facilitating insertion and removal. When inserting the connecting body 201 into the liquid reservoir, the connection is considered fully inserted until the limit portion 204 abuts the liquid reservoir. When removing the connecting body 201, force can be applied to the limit portion 204.

[0071] For example, the outer periphery of the connecting body 201 is provided with a sealing portion 205, which protrudes outward relative to the connecting body 201. When the connecting body 201 is inserted into the opening of the liquid reservoir, the sealing portion 205 abuts against the inner wall of the opening. The provision of the sealing portion 205 ensures a tight connection between the connecting body 201 and the liquid reservoir.

[0072] For example, the sealing portion 205 may be a circular ring structure.

[0073] For example, the outer periphery of the connecting body 201 is provided with a guide surface 2013, which is provided at one end of the connecting body 201 away from the connecting tube and is a conical surface. By providing the guide surface 2013, the connecting body 201 is facilitated to be inserted into the liquid reservoir.

[0074] For example, the limiting portion 204 may be cylindrical or prism-shaped, such as a hexagonal prism. The connecting body 201, the limiting portion 204 and the sealing portion 205 may be integrally formed to facilitate processing and production.

[0075] For example, referring to Figures 18 to 20, the connecting body 201 is a hollow structure, which saves materials and facilitates the processing of the mounting hole 2011 and the exhaust hole 2012. During the insertion of the liquid reservoir, the connecting body 201 is squeezed and elastically deformed. The hollow structure provides deformation space for elastic deformation, making it easier to insert the liquid reservoir. As shown in Figure 18, the hollow structure can be a conical groove 2014 provided inside the connecting body 201, and the opening of the conical groove 2014 is located at the bottom end of the connecting body 201; as shown in Figure 19, the hollow structure can be a cylindrical groove 2015 provided inside the connecting body 201, and the opening of the cylindrical groove 2015 is located at the bottom end of the connecting body 201; as shown in Figure 20, the hollow structure can be a cylindrical groove 2015 provided inside the connecting body 201, and the opening of the cylindrical groove 2015 is located at the top end of the connecting body 201.

[0076] Among them, any two or more of the limiting portion 204 , the sealing portion 205 , the guide surface 2013 and the hollow structure may exist at the same time.

[0077] The automatic liquid replenishment device and method for a sequencer provided in the embodiments of the present application simplify the structure, reduce costs, improve efficiency and facilitate maintenance.

[0078] The automatic liquid replenishment device and method for a sequencer proposed in the embodiments of the present application are connected through the cooperation of several liquid reservoirs, connectors, connecting tubes and peristaltic pumps. The connector is used to connect the liquid reservoir and the connecting tube, and the peristaltic pump is used to provide power for the flow of liquid. It has a simple structure, occupies a small space, has low production cost, is easy to disassemble and assemble, and is convenient to maintain. The liquid passes through the connector and the connecting tube to avoid residue and is easy to clean. It can automatically replenish a single solution and can also automatically replenish an instant mixed solution. It can automatically replenish and stop replenishing liquid at any time. It is easy to operate, reduces labor costs, improves work efficiency, and avoids errors.

[0079] The embodiments of the present application may be modified and varied, and all of these modifications and variations fall within the scope of the present application to be protected. The scope of protection claimed in this application is defined by the appended claims and their equivalents.

Claims

1. An automatic liquid replenishing device for a sequencer, comprising: A liquid reservoir, comprising at least a first liquid reservoir (11), a second liquid reservoir (12) and a third liquid reservoir (13); A connector, comprising at least a first connector (21), a second connector (22), a third connector (23) and a fourth connector (24), wherein the first connector (21) is arranged above the sequencer (100) and is in communication with a liquid inlet of the sequencing chip (101), the second connector (22) is arranged at the opening of the first liquid reservoir (11) and is in communication with the first liquid reservoir (11), the third connector (23) is arranged at the opening of the second liquid reservoir (12) and is in communication with the second liquid reservoir (12), and the fourth connector (24) is arranged at the opening of the third liquid reservoir (13) and is in communication with the third liquid reservoir (13); The connecting pipes at least comprise a first connecting pipe (31), a second connecting pipe (32), a third connecting pipe (33) and a fourth connecting pipe (34), wherein two ends of the first connecting pipe (31) are respectively connected to the first connector (21) and the second connector (22), two ends of the second connecting pipe (32) are respectively connected to the first connector (21) and the third connector (23), two ends of the third connecting pipe (33) are respectively connected to the third connector (23) and the fourth connector (24), and the fourth connecting pipe (34) is connected to the fourth connector (24); The peristaltic pump comprises at least a first peristaltic pump (41) and a second peristaltic pump (42), wherein the first peristaltic pump (41) is arranged on the path of the first connecting tube (31), and the second peristaltic pump (42) is arranged on the path of the fourth connecting tube (34).

2. The automatic liquid replenishing device for a sequencer according to claim 1, wherein: Each connector comprises a connecting body (201), the connecting body (201) being provided with at least one mounting hole (2011), each mounting hole (2011) being provided with a capillary tube (202), both ends of the capillary tube (202) respectively protruding from the connecting body (201), and the capillary tube (202) being connected with the corresponding liquid reservoir and the connecting tube.

3. The automatic liquid replenishing device for a sequencer according to claim 2, wherein: The capillary tube (202) is a stainless steel tube.

4. The automatic liquid replenishing device for a sequencer according to claim 2 or 3, wherein: Some of the connectors further include an extension tube (203), wherein the extension tube (203) is disposed in the corresponding liquid reservoir and is in communication with the capillary tube (202).

5. The automatic liquid replenishing device for a sequencer according to claim 4, wherein: The extension tube (203) is a silicone tube.

6. The automatic liquid replenishing device for a sequencer according to claim 2, wherein: A limiting portion (204) is provided at one end of the connection body (201) close to the corresponding connection pipe. The connection body (201) is inserted into the opening of the corresponding liquid reservoir. The limiting portion (204) is located outside the opening and abuts against the corresponding liquid reservoir.

7. The automatic liquid replenishing device for a sequencer according to claim 2, wherein: The outer ring of the connecting body (201) is provided with a sealing portion (205), and the sealing portion (205) protrudes outward relative to the connecting body (201). The connecting body (201) is inserted into the corresponding opening of the liquid reservoir, and the sealing portion (205) abuts against the inner wall of the opening.

8. The automatic liquid replenishing device for a sequencer according to claim 2, wherein: A guide surface (2013) is provided on the outer periphery of the connection body (201), and the guide surface (2013) is provided at an end of the connection body (201) away from the corresponding connection pipe, and the guide surface (2013) is a conical surface.

9. The automatic liquid replenishing device for a sequencer according to claim 2, wherein: The connecting body (201) is a hollow structure.

10. An automatic liquid replenishment method for a sequencer, applied to the automatic liquid replenishment device for a sequencer according to any one of claims 1 to 9, the method comprising: When a single solution is replenished, the first peristaltic pump (41) is started to squeeze the first connecting tube (31) alternately. As the gas in the first connecting tube (31) is emptied, the liquid in the first liquid reservoir (11) passes through the second connector (22) and the first connecting tube (31) to reach the first connector (21), and the liquid drips into the liquid inlet of the sequencing chip (101) in the form of droplets.

11. The automatic liquid replenishment method for a sequencer according to claim 10, further comprising: When replenishing the mixed solution immediately, the second peristaltic pump (42) rotates forward to draw the liquid in the second liquid reservoir (12) through the third connector (23), the third connecting tube (33) and the fourth connector (24) into the third liquid reservoir (13). After the liquid in the second liquid reservoir (12) is completely drawn out, the second peristaltic pump (42) rotates reversely to push the mixed solution in the third liquid reservoir (13) through the fourth connector (24), the third connecting tube (33), the second liquid reservoir (12), the third connector (23) and the second connecting tube (32) to reach the first connector (21), and the liquid drips into the liquid inlet of the sequencing chip (101) in the form of droplets.

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