Sequencing system and liquid inlet method

Abstract

The invention provides a sequencing system and a liquid inlet method, and the sequencing system comprises a sequencing chip, a reagent storage container, a liquid path system and a waste liquid storage container; a plurality of runners are arranged in the sequencing chip and are divided into a plurality of groups, and a plurality of runners are arranged in each group and are used for carrying out an amplification reaction and a sequencing reaction; the liquid path system comprises a plurality of multi-channel valves, a power pump and a multi-way valve, the number of the multi-channel valves is multiple, and the flow channel is communicated with the reagent storage container through the multi-channel valves; the plurality of multi-channel valves are in one-to-one correspondence with the plurality of flow channels in any group of flow channels, and the plurality of flow channels in each group of flow channels are respectively communicated to the corresponding multi-channel valves; the number of the power pumps is the same as the number of groups formed by the plurality of flow channels; the number of the multi-way valves is multiple, and two ports of each multi-way valve communicate with the corresponding flow channels and the corresponding power pump correspondingly. According to the technical scheme, the problem of reagent waste of the sequencing system is reduced, and resources are saved.

Description

Technical Field

[0001] The present application relates to the field of nucleic acid detection technology, and in particular to a sequencing system and a liquid injection method. Background Art

[0002] In the gene sequencing system, the pump provides power and distributes the liquid reagents through the multi-channel rotary valve so that the target liquid reagents enter the flow channel of the predetermined sequencing chip; different flow channels can enter and exit the same or different fluids at the same or different times, and the liquid path system controls the fluid in and out of the flow channel to sequentially perform fluid transport, waiting and reaction, etc. Other systems of the instrument also perform their respective tasks in the above order.

[0003] The reaction of the reagents in the flow channel only occurs in a very thin area, which is much thinner than the thickness of the flow channel. The effective components of the reagents participating in the reaction are usually less than 10%, or even less than 1%. The reagents participating in the reaction in the flow channel are discharged to the waste liquid, which is a serious waste problem. Summary of the invention

[0004] The present application provides a sequencing system and a liquid injection method to reduce the problem of reagent waste in the sequencing system and save resources.

[0005] In a first aspect, the present application provides a sequencing system, including a sequencing chip, a reagent storage container, a liquid path system, and a waste liquid storage container;

[0006] The sequencing chip is provided with a plurality of flow channels, and the plurality of flow channels are divided into a plurality of groups, and each group is provided with a plurality of channels for amplification reaction and sequencing reaction;

[0007] The fluid path system comprises a multi-channel valve, a power pump and a multi-way valve, wherein there are multiple multi-channel valves, and the flow channel is connected to the reagent storage container through the multi-channel valve;

[0008] The plurality of multi-channel valves correspond one-to-one to the plurality of flow channels in any group of the flow channels, and the plurality of flow channels in each group of the flow channels are respectively connected to the corresponding multi-channel valves;

[0009] The number of the power pumps is the same as the number of groups formed by the plurality of flow channels, and corresponds one-to-one to the plurality of groups of the flow channels;

[0010] The multi-way valves are provided in plurality and correspond one-to-one with the plurality of flow channels, and two ports of each multi-way valve are respectively connected with the corresponding flow channel and the power pump;

[0011] Each of the multi-way valves is also provided with at least one port communicating with the waste liquid storage container.

[0012] In a specific possible implementation scheme, each of the multi-channel valves is provided with a corresponding multi-port, and one of the interfaces of the multi-port is communicated with the common port of the multi-channel valve;

[0013] The other interfaces of the multi-channel valve are in one-to-one communication with the plurality of flow channels corresponding to the multi-channel valve.

[0014] In a specific implementation scheme, there are four flow channels in total, with two channels forming a group.

[0015] In a specific embodiment, the multi-port is directly connected to the common port of the multi-channel valve.

[0016] In a second aspect, the present application further provides a method for liquid injection into a sequencing system, which is implemented based on any of the sequencing systems described above, and the method comprises the following steps:

[0017] Step S1: simultaneously inputting reagent a into a plurality of flow channels α in one group of the flow channels;

[0018] Step S2: The reagent a in any of the flow channels α is driven by a corresponding power pump, flows through the multi-channels connected to the flow channel α, and is transported to another flow channel β in a different group connected to the same multi-channel;

[0019] Step S3: injecting reagent b into the flow channel α again.

[0020] In a specific implementation scheme, specifically:

[0021] Inputting reagent a into all flow channels α of the same group;

[0022] After the reagent a has completed its reaction in the flow channel α, the reacted reagent a is transported to each of the flow channels β in another group of the flow channels;

[0023] and replenishing unreacted reagent a into each of the flow channels β;

[0024] While reagent a is reacting in the flow channel β, reagent b is reacting in the flow channel α.

[0025] In a specific embodiment, after reagent a is in the flow channel β and reagent b is reacted in the flow channel α, the reacted reagent b is transported from the flow channel α to the flow channel β, and at the same time, the reagent a in the flow channel β is transported to other flow channels or a waste liquid storage container;

[0026] The reagent c is transported into the flow channel α, and the unreacted reagent b is added into the flow channel β.

[0027] In a specific implementation scheme, specifically:

[0028] Delivering reagent a into one of the flow channels α1 in a group of flow channels;

[0029] The reagent a in the flow channel α1 flows through the connected multi-channel and is transported to one of the flow channels β1 in another group of flow channels;

[0030] Replenishing reagent a into the flow channel β1, and delivering reagent a or reagent b into the flow channel α1;

[0031] The reagents of corresponding types react in the flow channel α1 and the flow channel β1;

[0032] The flow channel α1 and the flow channel β1 are connected to the same multi-channel.

[0033] In a specific embodiment, the reagent b is delivered into one of the flow channels α2 in a group of flow channels;

[0034] The reagent b in the flow channel α2 flows through the connected multi-channel and is transported to one of the flow channels β2 in another group of flow channels;

[0035] Replenishing reagent b into the flow channel β2, and delivering reagent b or reagent a or reagent c into the flow channel α2;

[0036] The reagents of corresponding types react in the flow channel α2 and the flow channel β2;

[0037] Wherein, the flow channel α2 and the flow channel β2 are connected to the same multi-channel.

[0038] In a specific embodiment, the reagent is delivered into the flow channel to complete the liquid feeding process, and after the reaction in each of the flow channels is completed, an image is collected by an optical detection system;

[0039] After the image acquisition process is completed, the reagent is re-delivered to the corresponding flow channel, and a continuous cycle of liquid injection-reaction-image acquisition is maintained;

[0040] The optical detection system performs a process of collecting images of the flow channel α1 and the flow channel β1, and a process of collecting images of the flow channel α2 and the flow channel β2, respectively.

[0041] In a specific implementation scheme, in step S1, the multi-channel valve corresponding to the flow channel α is connected to the reagent storage container, and when the multi-channel valve corresponding to the flow channel α is adjusted, the flow channel α is connected to the corresponding power pump, and the power pump sucks liquid to deliver the reagent to the flow channel α;

[0042] In step S2, the common ports of the multi-channel valves corresponding to the flow channel α and the flow channel β are connected to the blocking port, the multi-way valve corresponding to the flow channel β is adjusted so that the flow channel β is connected to the waste liquid storage container, and the power pump pushes back to transport the reagent from the flow channel α to the flow channel β. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 An overall schematic diagram of the sequencing system provided in the embodiments of the present application;

[0044] Figure 2 A schematic diagram of the relative relationship between the fluid circuit system and the flow channel and other components provided in the embodiment of the present application;

[0045] Figure 3 A schematic diagram of the structure of the liquid circuit system provided in an embodiment of the present application;

[0046] Figure 4 A schematic diagram of a reagent transmission path in the first step of a liquid feeding method provided in an embodiment of the present application;

[0047] Figure 5 A schematic diagram of a reagent transmission path in the second step of a liquid feeding method provided in an embodiment of the present application;

[0048] Figure 6 A schematic diagram of a transmission path for delivering a reagent to a portion of a flow channel in a liquid feeding method provided in another embodiment of the present application;

[0049] Figure 7 A schematic diagram of a transmission path for delivering a reagent to another portion of a flow channel in a liquid injection method provided in another embodiment of the present application. DETAILED DESCRIPTION

[0050] In order to make the objectives, technical solutions and advantages of the present application clearer, the present application will be further described in detail below in conjunction with the accompanying drawings.

[0051] It should be noted that, unless otherwise defined, the technical terms or scientific terms used in one or more embodiments of this specification should be understood by people with ordinary skills in the field to which this disclosure belongs. The "first", "second" and similar words used in one or more embodiments of this specification do not indicate any order, quantity or importance, but are only used to distinguish different components. "Include" or "comprise" and similar words mean that the elements or objects appearing before the word cover the elements or objects listed after the word and their equivalents, without excluding other elements or objects. "Connect" or "connected" and similar words are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect. "Up", "down", "left", "right" and the like are only used to indicate relative positional relationships. When the absolute position of the described object changes, the relative positional relationship may also change accordingly.

[0052] To facilitate understanding of the sequencing system and liquid feeding method provided in the embodiment of the present application, its application scenario is first described. The sequencing system is used for gene sequencing work, including a sequencing chip, a chip platform, a reagent storage container, a liquid circuit system, etc., wherein the liquid circuit system is used for the transportation of sample libraries and reagents, transporting reagents to the sequencing chip for reaction, and transporting the reacted reagents to the waste liquid. Among them, amplification reaction and sequencing reaction occur in the sequencing chip, and image acquisition is performed by an image acquisition system to achieve gene sequencing. Exemplarily, a plurality of mutually isolated flow channels are provided in the sequencing chip, and the above reaction process is carried out in the flow channel, and the reaction only exists in a very thin area, the thickness of this area is much smaller than the flow channel thickness, that is, only part of the reagents transported to each flow channel participate in the reaction, and the remaining part is directly discharged to the waste liquid for collection after the reaction is completed; the effective ingredients of the reagents participating in the reaction are usually within 10%, or even within 1%, and the remaining reagents are all discharged to the waste liquid for centralized treatment, resulting in a more serious waste problem; and the cost of some of the reagents is high, which will also cause the problem of increased overall cost.

[0053] To this end, the present application provides a sequencing system and a liquid injection method to reduce reagent waste during sequencing, save resources, and reduce costs. The following is a detailed description of the system and the embodiments in conjunction with the specific drawings.

[0054] First reference Figure 1 , Figure 1The overall schematic diagram of the sequencing system is shown. The sequencing system includes a sequencing chip 10, a chip platform 20, a reagent storage container 30, a liquid circuit system 40, an optical detection system 50, a computer system 60 and a waste liquid storage container 70. Among them: the sequencing chip 10 is configured to provide a reaction area for amplification reaction and sequencing reaction; the chip platform 20 is configured to fix and support the sequencing chip 10; the reagent storage container 30 is configured to store one or more mixed sample libraries, one or more reagents; the liquid circuit system 40 is configured to controllably transport one or more mixed sample libraries, one or more reagents from the reagent storage container 30 to the sequencing chip 10, so as to perform amplification reaction and sequencing reaction in the sequencing chip 10, and controllably transport the waste liquid after the reaction from the sequencing chip 10 to the waste liquid storage container 70.

[0055] The optical detection system 50 is configured to excite and collect fluorescent signals during the sequencing reaction and generate fluorescent images based on the fluorescent signals; the computer system 60 is configured to obtain the fluorescent images from the optical detection system 50 and identify the base sequence of the sample library based on the fluorescent images; the waste liquid storage container 70 is configured to store the waste liquid generated after the reaction.

[0056] refer to Figure 2 , shows the connection relationship between the liquid circuit system 40 and the sequencing chip 10, the reagent storage container 30 and the waste liquid storage container 70, wherein the sequencing chip 10 is provided with multiple flow channels 11, and the multiple flow channels 101 are divided into multiple groups, each group includes multiple channels, as the place for diffusion reaction and sequencing reaction. The liquid circuit system 40 includes a multi-channel valve 41, a power pump 42 and a multi-way valve 43, and also includes pipelines for connecting various structures. In the embodiment of the present application, for the convenience of description and understanding, each section of the pipeline is no longer defined separately, but only defined in the way that the corresponding components are connected or connected. In actual design, the connection between components is realized through the corresponding pipeline structure.

[0057] There are multiple multi-channel valves 41, which are arranged between the sequencing chip 10 and the reagent storage container 30, and the multiple flow channels 11 are connected to the reagent storage container 30 through the multiple multi-channel valves 41. Specifically, the number of multi-channel valves 41 is the same as the number of each group of flow channels 101, that is, the multiple multi-channel valves 41 correspond to the multiple flow channels 11 in any group of flow channels 11, and the multiple flow channels 11 included in each group are connected to the multiple multi-channel valves 41 in a one-to-one correspondence.

[0058] Specifically, the multi-channel valve 41 is provided with a common port 411, a blocking port 412 and a plurality of branch ports 413, wherein the common port 411 can be selectively connected to the blocking port 412 and the plurality of branch ports 413; the multi-channel valve 41 is a structure conventionally adopted in the art, and the specific implementation of its functions will not be repeated in the embodiments of the present application.

[0059] Multiple flow channels 11 connected to the same multi-channel valve 41 are connected in parallel to the common port 411, and the branch port 413 is connected to the reagent storage container 30 to transport the reagent to the flow channel 11 through the multi-channel valve 41; at least one of the multiple branch ports 413 is connected to the waste liquid storage container 70 for discharging waste liquid.

[0060] The number of power pumps 42 is set to be the same as the number of groups formed by the multiple flow channels 11, and corresponds one-to-one to the multiple groups of flow channels 11. The multiple flow channels 11 in the same group are connected to the corresponding power pumps 42. During the process of liquid filling and discharge, the power pumps 42 provide power for transporting reagents.

[0061] There are multiple multi-way valves 43, corresponding to the multiple flow channels 11 one by one, each multi-way valve 43 is connected between the corresponding flow channel 11 and the power pump 42, and two of the ports of the multi-way valve 43 are respectively connected to the corresponding flow channel 11 and the power pump 42. In addition, each multi-way valve 43 is also provided with at least one port connected to the waste liquid storage container 70. Any two of the multiple ports of the multi-way valve 43 can be selectively connected. In the embodiment of the present application, the multi-way valve 43 takes a three-way valve as an example. Of course, in other embodiments, a four-way valve, a five-way valve, etc. can also be selected.

[0062] In addition, the liquid circuit system 40 also includes multiple multi-channels 44, and the multiple multi-channels 44 are arranged in a one-to-one correspondence with the multiple multi-channel valves 41. One of the interfaces of each multi-channel 44 is connected to the common port of the corresponding multi-channel valve 41, and the other interfaces are connected to the multiple flow channels 11 corresponding to the multi-channel valve 41 in a one-to-one correspondence. Exemplarily, the multi-channel 41 is connected to the flow channel 11 through a connecting pipe, and the multiple flow channels 11 are connected in parallel through the set multi-channel 44 and are connected to the corresponding multi-channel valve 41.

[0063] refer to Figure 3 , and combined with reference Figure 2 , taking the example that there are four flow channels 11, they are divided into two groups, each with two; correspondingly, the multi-channel valve 41 and the power pump 42 are each provided with two, and the multi-way valve 43 is provided with four; for the convenience of description, the two groups of flow channels 11 are respectively group α and group β, and in the case of more groups, they can also be named as γ, δ, etc. Specifically, the four flow channels 11 are flow channel α1, flow channel α2, flow channel β1, and flow channel β2, wherein flow channel α1 and flow channel β1 are connected to the multi-channel valve 41 in parallel with the same multi-way 44, and flow channel α2 and flow channel β2 are connected to the multi-channel valve 41 in parallel with the same multi-way 44. When each group has three or more flow channels 11, flow channel α3, flow channel α4, etc. are used to distinguish them. The two multi-channel valves 41 are respectively multi-channel valve A and multi-channel valve B, the two power pumps 42 are respectively power pump A and power pump B, and the four multi-way valves 43 are respectively multi-way valve a, multi-way valve b, multi-way valve c and multi-way valve d.

[0064] The specific liquid injection process is as follows: the multi-channel valve A and the multi-channel valve B are connected to the reagent a, and the multi-channel valve a and the multi-channel valve b are adjusted so that the flow channel α1 and the flow channel α2 are connected to the power pump A, and the power pump A sucks liquid and delivers the reagent a to the flow channel α1 and the flow channel α2, and the reagent a reacts in the two flow channels 11. After the reaction is completed, the multi-channel valve A and the multi-channel valve B are adjusted to connect the common port 411 and the blocking port 412, and the power pump A pushes back to deliver the reacted reagent a in the flow channel α1 and the flow channel α2 to the flow channel β1 and the flow channel β2 of another group through the corresponding multi-port 44.

[0065] Afterwards, multi-channel valve A and multi-channel valve B are connected to reagent b. By adjusting the corresponding multi-way valve a and multi-way valve b, and power pump A, reagent b is transported to flow channel α1 and flow channel α2. The specific process can refer to the process of transporting reagent a to flow channel α1 and flow channel α2.

[0066] It should be noted that the reagent a and reagent b defined in the embodiments of the present application, or other subsequent reagents c, reagent d, etc., may be reagents of the same type or reagents of different types.

[0067] In the above process, part of the unreacted reagent a can be added to the flow channel β1 and the flow channel β2 as needed to meet the requirements of the reaction of reagent a. For example, the multi-channel valve A and the multi-channel valve B are connected to the reagent a, and the multi-way valve c and the multi-way valve d corresponding to the flow channel β1 and the flow channel β2 are adjusted so that the flow channel β1 and the flow channel β2 are connected to the power pump B, and the power pump B sucks liquid and delivers the reagent a to the flow channel β1 and the flow channel β2. It can ensure that the amount of the reagent meets the standard for the normal reaction, avoiding the problem of deviation in the results due to insufficient amount of the reagent.

[0068] By setting up the liquid path system 40, the reagents can be reused, which reduces the problem of the reagents being directly discharged as waste liquid after reacting once, causing waste; and in the process of reusing the reagents, the reagents in the pipeline between the multi-channel 44 and the flow channel 11 can also be recovered and reused, further reducing the actual waste problem.

[0069] In some embodiments, multi-way 44 is directly connected to common port 411 of multi-channel valve 41 to reduce the amount of reagent that can be retained between multi-way 44 and multi-channel valve 41. In the case of reusing reagents, the amount of reagents that are not reused can be further reduced. In other embodiments, the length of the connecting pipeline between multi-way 44 and multi-channel valve 41 can also be shortened as much as possible to reduce the amount of the reagent that is not reused.

[0070] In addition, for the above-mentioned process of transporting reagent a and reagent b to different flow channels 11, in the actual liquid feeding process, cleaning liquid, buffer solution, etc. can also be interspersed between the transport of different reagents to clean the flow channel 11 to avoid contamination between different reagents. Exemplarily, after reagent a is transferred from flow channel α1 and flow channel α2 to flow channel β1 and flow channel β2, the cleaning liquid is transported to flow channel α1 and flow channel α2 for cleaning. After cleaning, the cleaning liquid is discharged from the corresponding multi-channel valve 41 to the waste liquid storage container 70, and then the subsequent reagent b is transported. In the embodiments of the present application, such processes are no longer described in detail. In actual operation, different types of reagents can be introduced according to actual needs.

[0071] When different types of reagents are transported to the flow channel 11, the multi-channel valve 41 is required to be connected to different reagent storage containers 30. In some embodiments, a pipeline is connected to one end of the multi-channel valve 41 facing the reagent storage container 30. When the transported reagent needs to be replaced, the pipeline is connected to the corresponding reagent storage container 30.

[0072] In other embodiments, after multiple multi-channel valves 41 are connected in parallel through pipelines, they are connected to different reagent storage containers 30 through solenoid valves. Specifically, the solenoid valve is provided with multiple ports, and the multiple ports can be selectively connected at will. After multiple multi-channel valves 41 are connected in parallel, they are connected to one of the ports of the solenoid valve, and the other multiple ports of the solenoid valve are respectively connected to different reagent storage containers 30; the multiple ports of the solenoid valve can be selectively connected, and when different reagents need to be transported, different ports of the solenoid valve can be adjusted to connect with the multi-channel valve 41.

[0073] The embodiment of the present application also provides a liquid injection method for a sequencing system, which is implemented based on the above sequencing system. Specifically, the method includes the following steps:

[0074] Step S1: delivering reagent a to a plurality of flow channels α in one group of flow channels 11 simultaneously;

[0075] Step S2: For all flow channels α input with reagent a, reagent a in any flow channel α is driven by the corresponding power pump 42, flows through the multi-channel 44 connected to the corresponding flow channel α, and is transported to another flow channel β of a different group connected to the multi-channel;

[0076] Step S3: Afterwards, the reagent b is input into the flow channel α again, and according to actual needs, a certain amount of unreacted reagent a is supplemented and transported into the flow channel β.

[0077] It should be explained that the flow channel α and the flow channel β defined in the embodiment of the present application may refer to one of the flow channels 11 in the same group or multiple flow channels 11 in the same group; taking two groups of flow channels 11 with two flow channels in each group as an example, the four flow channels 11 are flow channel α1, flow channel α2, flow channel β1, and flow channel β2; then the flow channel α is flow channel α1 and / or flow channel α2, and the flow channel β is flow channel β1 and / or flow channel β2.

[0078] In step S1, the common port 411 of the multi-channel valve 41 corresponding to the flow channel α is connected to the reagent storage container 30, and the multi-way valve 43 corresponding to the flow channel α is adjusted so that the flow channel α is connected to the corresponding power pump 42, and the power pump 42 sucks liquid to deliver the reagent to the flow channel α.

[0079] In step S2, the common ports 411 of the multi-channel valves 41 corresponding to the flow channel α and the flow channel β are connected to the blocking port 412, the multi-way valve 43 corresponding to the flow channel β is adjusted to connect the flow channel β with the waste liquid storage container 70, and the power pump 42 is pushed back to transport the reagent from the flow channel α to the flow channel β.

[0080] The reagent a is simultaneously delivered to several flow channels α in a group of flow channels 11 , which may be delivered to all the flow channels 11 of the group at the same time, or delivered to one of the flow channels 11 .

[0081] refer to Figure 4 and Figure 5 In some embodiments, the above-mentioned simultaneous delivery of reagent a to several flow channels α in one group of flow channels 11 is to simultaneously input reagent a into all flow channels 11 of the group, specifically, including the following steps.

[0082] Reagent a is transported to all flow channels α of the same group.

[0083] After reagent a completes the reaction in flow channel α, the reacted reagent a is transported to each flow channel β of another group of flow channels 11 through the corresponding multi-channel 44; wherein, flow channel α corresponds to flow channel β one by one, and the corresponding two flow channels 11 are connected through the multi-channel 44 to realize the transmission of the reagent.

[0084] in, Figure 4 The schematic diagram of the reagent transmission path when the reagent is transported into the flow channel α is shown as an example. Figure 5 The schematic diagram of the reagent transmission path in the process of transporting the reagent from the flow channel α to the flow channel β is exemplarily shown.

[0085] Reagent b is delivered to all flow channels α of the same group, and a certain amount of unreacted reagent a is added to flow channel β according to actual needs.

[0086] As described above, while reagent a is reacting in flow channel β, reagent b is reacting in flow channel α. After the reaction is completed, other reagents are transported therein, and then other subsequent reaction processes are carried out.

[0087] Exemplarily, the reacted reagent b is transported from the flow channel α to the flow channel β, and at the same time, the reagent a in the flow channel β is transported to the waste liquid storage container 70. If the flow channel 11 is provided with other groups, such as the flow channel γ, the flow channel δ, etc., the reagent a in the flow channel β can also be transported to other flow channels 11 for reuse, and then the reagent b in the flow channel α can be transported to the flow channel β for reuse.

[0088] Afterwards, new reagent c is transported into the flow channel α, and a certain amount of unreacted reagent b is added to the flow channel β as needed.

[0089] Taking the example of setting four flow channels 11 in the above-mentioned sequencing system as an example, the specific steps of the liquid injection method are as follows: the common port 411 of the multi-channel valve A and the multi-channel valve B are connected to the reagent storage container 30 for storing reagent a, and the multi-way valve a and the multi-way valve b are adjusted to the state where the corresponding flow channel α1 and the flow channel α2 are connected to the corresponding power pump A, and the power pump A sucks liquid to transport the reagent a to the flow channel α and the flow channel β.

[0090] Reagent a reacts in flow channel α1 and flow channel α2. After the reaction is completed, the common port 411 of multi-channel valve A and multi-channel valve B is connected to the blocking port 412, and the multi-way valve c and the multi-way valve d are adjusted to the state where the corresponding flow channel β1 and flow channel β2 are connected to the waste liquid storage container 70; the power pump A performs a back-pushing action to transport the reagent a in the flow channel α1 and flow channel α2 to the flow channel β1 and flow channel β2.

[0091] Adjust the multi-channel valve A and the multi-channel valve B again so that the common port 411 is connected to the reagent b, adjust the multi-way valve a, the multi-way valve b and the power pump A to operate, and deliver the reagent b to the flow channel α1 and the flow channel α2; in the process of delivering the reagent b, the adjustment of the multi-way valve a, the multi-way valve b and the power pump A can refer to the process of delivering the reagent a to the flow channel α1 and the flow channel α2.

[0092] If it is necessary to add a certain amount of unreacted reagent a into the flow channel β1 and the flow channel β2, adjust the multi-way valve c and the multi-way valve d so that the flow channel β1 and the flow channel β2 are connected to the power pump B. Correspondingly, the common port 411 of the multi-channel valve A and the multi-channel valve B is connected to the reagent a, and the power pump B sucks liquid to transport part of the unreacted reagent a into the flow channel β1 and the flow channel β2.

[0093] Reagent a and reagent b react in the corresponding flow channel 11. After the reaction is completed, the common port 411 of the multi-channel valve A and the multi-channel valve B are connected to their respective blocking ports 412. The multi-way valve c and the multi-way valve d are adjusted so that the flow channels β1 and β2 are connected to the waste liquid storage container 70. The power pump A performs a back-pushing action to push the reagent b in the flow channels α1 and α2 into the flow channels β1 and β2, and at the same time, transport the reagent a from the flow channels β1 and β2 to the waste liquid storage container 70 for recovery.

[0094] Afterwards, reagent c is delivered to flow channel α1 and flow channel α2 by adjusting the multi-channel valves 41, power pump 42 and multi-way valve 43, and a certain amount of unreacted reagent b is added to flow channel β1 and flow channel β2 as needed. Subsequently, reagent c, reagent d, etc. can be reused according to the above method to reduce the waste of reagents during sequencing.

[0095] refer to Figure 6 and Figure 7 In other embodiments, the above-mentioned simultaneous delivery of reagent a to several flow channels α in one group of flow channels 11 is to deliver reagent a to one flow channel α1 in one group of flow channels 11, specifically, including the following steps:

[0096] Delivering reagent a into one of the flow channels α1 in a group of flow channels 11;

[0097] The reagent a in the control flow channel α1 flows through the connected multi-channel 44 and is transported to one of the flow channels β1 in another group of flow channels 11;

[0098] A certain amount of unreacted reagent a is added to the flow channel β1, and reagent a or reagent b is transported into the flow channel α1. That is, the same or different type of reagent as that in the flow channel β1 can be transported into the flow channel α1. In the embodiment of the present application, the transport of reagent b is taken as an example.

[0099] The above-mentioned corresponding types of reagents react in the flow channel α1 and the flow channel β1, wherein the flow channel α1 and the flow channel β1 are connected to the same multi-channel 44.

[0100] Figure 6 The schematic diagram of the reagent transmission path in the process of transporting the reagent into the flow channel α1 and the flow channel β1 is exemplarily shown, wherein I and II represent the order of the corresponding transmission paths; Figure 7 The schematic diagram of the reagent transmission path during the process of transporting the reagent into the flow channel α2 and the flow channel β2 is exemplarily shown, wherein I and II represent the order of the corresponding transmission paths.

[0101] While the reagents react in the flow channel α1 and the flow channel β1, liquid is introduced into the flow channel α2 and the flow channel β2. Specifically, the following steps are included: a reagent b is transported into one of the flow channels α2 in a group of flow channels 11;

[0102] The reagent b in the flow channel α2 flows through the connected multi-channel 44 and is transported to one of the flow channels β2 in another group of flow channels 11;

[0103] Reagent b is added to flow channel β2, and reagent b, reagent a, or reagent c is delivered to flow channel α2. That is, the type of reagent delivered can be determined according to actual needs, and there is no restriction on the type of reagent;

[0104] The above corresponding types of reagents react in the flow channel α2 and the flow channel β2.

[0105] As described above, the process of transporting reagents into the flow channel 11 includes two parts: first, transporting reagents into the flow channel α1 and the flow channel β1, and reagents of corresponding types react in the flow channel α1 and the flow channel β1; second, transporting reagents into the flow channel α2 and the flow channel β2, and reagents of corresponding types react in the flow channel α2 and the flow channel β2.

[0106] The above two processes are carried out alternately, that is, while the reagent reacts in the flow channel α1 and the flow channel β1, liquid is introduced into the flow channel α2 and the flow channel β2; while the reagent reacts in the flow channel α2 and the flow channel β2, liquid is introduced into the flow channel α1 and the flow channel β1.

[0107] In addition, during the reagent reaction, image acquisition is performed through the optical detection system 50, so that the reaction and image acquisition processes are performed alternately with the liquid injection process to improve efficiency. In addition, considering that the optical detection system 50 has a certain volume and the spacing between the flow channels 11 cannot be too large to ensure that the sequencing chip 10 has a smaller size, the optical detection system 50 performs image acquisition on the flow channels α1 and β1, and the flow channels α2 and β2 with a certain spacing, thereby improving the liquid injection reaction efficiency and the image acquisition efficiency.

[0108] After the reaction process in flow channel α1 and flow channel β1 is completed, the corresponding reaction starts in flow channel α2 and flow channel β2. At this time, new reagents are transported to flow channel α1 and flow channel β1. At this time, the reagents in flow channel α1 can be transported to flow channel β1 for reuse, that is, the reagents in flow channel β1 are transported to waste liquid storage container 70, the reagents in flow channel α1 are transported to flow channel β1, and new reagents are transported to flow channel α1. The reagents can also be reused in the corresponding liquid inlet method to reduce waste problems.

[0109] In other embodiments of the liquid injection method, reagent a can also be delivered to the flow channel α1 and the flow channel β1 at the same time. Specifically, the common port 411 of the multi-channel valve A is connected to the reagent storage container 30 for storing reagent a, and the multi-way valve a and the multi-way valve c are adjusted to the state where the corresponding flow channel α1 and the flow channel β1 are connected to the corresponding power pump A and the power pump B, and the power pump A and the power pump B suck the liquid to deliver the reagent a to the flow channel α1 and the flow channel β1.

[0110] During the reaction of reagent a in flow channel α1 and flow channel β1, images of the inside of flow channel α1 and flow channel β1 are collected by optical detection system 50. At the same time, reagent b is delivered to flow channel α2 and flow channel β2. The specific process is as follows: the common port 411 of multi-channel valve B is connected to the reagent storage container 30 storing reagent b, multi-way valve b and multi-way valve d are adjusted to the state where the corresponding flow channel α2 and flow channel β2 are connected to the corresponding power pump A and power pump B, and multi-way valve a and multi-way valve c are adjusted to the state where the corresponding flow channel α1 and flow channel β1 are connected to the waste liquid storage container 70; power pump A and power pump B suck liquid and deliver reagent to flow channel α2 and flow channel β2.

[0111] After the reagents are transported into the flow channel α2 and the flow channel β2, the reagents react in the flow channel α2 and the flow channel β2, and an image is captured by the optical detection system 50 after the reaction is completed.

[0112] Specifically, during the working process, the reagent is transported into the flow channel 11 to complete the liquid filling process, and after the reaction in each flow channel 11 is completed, the image is collected by the optical detection system 50; after the image collection process is completed, the reagent is re-transported to the corresponding flow channel 11, and during the working process within a certain period of time, the liquid filling-reaction-image collection is maintained continuously; and the optical detection system 50 performs the image collection process for the flow channel α1 and the flow channel β1, and the image collection process for the flow channel α2 and the flow channel β2, respectively.

[0113] That is, during the working process, the process of liquid inlet-reaction-image collection for flow channel α1 and flow channel β1, and the process of liquid inlet-reaction-image collection for flow channel α2 and flow channel β2 are all carried out in a continuous cycle, that is, the liquid inlet process continues after the image collection is completed; and the image collection process of flow channel α1 and flow channel β1 is carried out separately from the image collection process of flow channel α2 and flow channel β2, and is staggered in time. For example, in the process of image collection for flow channel α2 and flow channel β2, flow channel α1 and flow channel β1 are in the process of liquid inlet or reaction. It ensures that multiple flow channels 11 are in continuous operation, avoiding the situation where different processes need to wait for a certain time before switching to another process, thereby improving work efficiency.

[0114] According to the above method, the processes of delivering reagents to flow channels α1 and β1, and delivering reagents to flow channels α2 and β2 are performed alternately, so that the entire sequencing system, the process of delivering reagents, reacting with reagents, and collecting images can be carried out continuously, thereby improving efficiency.

[0115] One or more embodiments of this specification are intended to cover all such substitutions, modifications and variations that fall within the broad scope of the appended claims. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of one or more embodiments of this specification should be included in the scope of protection of this disclosure.

[0116] The above are only specific implementations of the present application, but the protection scope of the present application is not limited thereto. Any technician familiar with the technical field can easily think of changes or substitutions within the technical scope disclosed in the present application, which should be included in the protection scope of the present application. Therefore, the protection scope of the present application should be based on the protection scope of the claims.

Claims

1. A sequencing system, characterized in that: Including sequencing chip, reagent storage container, liquid system and waste liquid storage container; The sequencing chip is provided with a plurality of flow channels, and the plurality of flow channels are divided into a plurality of groups, and each group is provided with a plurality of channels for amplification reaction and sequencing reaction; The fluid path system includes a multi-channel valve, a power pump and a multi-way valve, wherein there are multiple multi-channel valves, and the flow channel is connected to the reagent storage container through the multi-channel valve; The plurality of multi-channel valves correspond one-to-one to the plurality of flow channels in any group of the flow channels, and the plurality of flow channels in each group of the flow channels are respectively connected to the corresponding multi-channel valves; The number of the power pumps is the same as the number of groups formed by the plurality of flow channels, and corresponds one-to-one to the plurality of groups of the flow channels; The multi-way valves are provided in plurality and correspond one-to-one with the plurality of flow channels, and two ports of each multi-way valve are respectively connected with the corresponding flow channel and the power pump; Each of the multi-way valves is also provided with at least one port communicating with the waste liquid storage container.

2. The sequencing system according to claim 1, characterized in that Each of the multi-channel valves is correspondingly provided with a multi-port, and one of the interfaces of the multi-port is communicated with the common port of the multi-channel valve; The other interfaces of the multi-channel valve are in one-to-one communication with the plurality of flow channels corresponding to the multi-channel valve.

3. The sequencing system according to claim 2, characterized in that There are four flow channels in total, with two of them forming a group.

4. The sequencing system according to claim 2, characterized in that The multi-port is directly connected to the common port of the multi-channel valve.

5. A method for liquid injection in a sequencing system, characterized in that: Based on the sequencing system according to any one of claims 1 to 4, the method comprises the following steps: Step S1: simultaneously inputting reagent a into a plurality of flow channels α in one group of the flow channels; Step S2: The reagent a in any of the flow channels α is driven by a corresponding power pump, flows through the multi-channels connected to the flow channel α, and is transported to another flow channel β in a different group connected to the same multi-channel; Step S3: injecting reagent b into the flow channel α again.

6. The liquid inlet method of the sequencing system according to claim 5, characterized in that: Specifically: Inputting reagent a into all flow channels α of the same group; After the reagent a has completed its reaction in the flow channel α, the reacted reagent a is transported to each of the flow channels β in another group of the flow channels; and replenishing unreacted reagent a into each of the flow channels β; While reagent a is reacting in the flow channel β, reagent b is reacting in the flow channel α.

7. The liquid inlet method of the sequencing system according to claim 6, characterized in that: After reagent a and reagent b have finished reacting in the flow channel α, the reacted reagent b is transported from the flow channel α to the flow channel β, and at the same time, the reagent a in the flow channel β is transported to other flow channels or a waste liquid storage container; The reagent c is transported into the flow channel α, and the unreacted reagent b is added into the flow channel β.

8. The liquid inlet method of the sequencing system according to claim 5, characterized in that: Specifically: Delivering reagent a into one of the flow channels α1 in a group of flow channels; The reagent a in the flow channel α1 flows through the connected multi-channel and is transported to one of the flow channels β1 in another group of flow channels; Replenishing reagent a into the flow channel β1, and delivering reagent a or reagent b into the flow channel α1; The reagents of corresponding types react in the flow channel α1 and the flow channel β1; The flow channel α1 and the flow channel β1 are connected to the same multi-channel.

9. The liquid inlet method of the sequencing system according to claim 8, characterized in that: Delivering reagent b into one of the flow channels α2 in a group of flow channels; The reagent b in the flow channel α2 flows through the connected multi-channel and is transported to one of the flow channels β2 in another group of flow channels; Replenishing reagent b into the flow channel β2, and delivering reagent b or reagent a or reagent c into the flow channel α2; The reagents of corresponding types react in the flow channel α2 and the flow channel β2; Wherein, the flow channel α2 and the flow channel β2 are connected to the same multi-channel.

10. The liquid inlet method of the sequencing system according to claim 9, characterized in that: The reagent is transported into the flow channel to complete the liquid feeding process, and after the reaction in each of the flow channels is completed, an image is collected by an optical detection system; After the image acquisition process is completed, the reagent is re-delivered to the corresponding flow channel, and a continuous cycle of liquid injection-reaction-image acquisition is maintained; The optical detection system performs a process of collecting images of the flow channel α1 and the flow channel β1, and a process of collecting images of the flow channel α2 and the flow channel β2, respectively.

Images