Liquid handling system and method for DNA synthesis, DNA synthesis system

Abstract

The application discloses a liquid path supply system and method for DNA synthesis and a DNA synthesis system. The liquid path supply system comprises a reaction reagent supply system, a switching system and a cleaning source supply system. The cleaning source supply system comprises a reaction reagent cleaning system matched with the reaction reagent supply system and a valve island cleaning system matched with the switching system. The switching system comprises a multi-return valve island and a reagent switching assembly. The valve island has an outlet and a plurality of inlets on the side surface, and is internally provided with an input branch channel, a conveying branch channel and an output main channel. One end of the input branch channel is communicated with the corresponding inlet, and the other end is connected with the switching system. The conveying branch channel is connected with the outlet of the switching system. The outlets of all the conveying branch channels converge in the same intersection area, and the intersection area is communicated with the inlet of the output main channel. The outlet of the output main channel is communicated with a common outlet. The method can realize pollution-free switching of different reagents into the corresponding common pipeline of the closed reaction cavity.

Description

TECHNICAL FIELD

[0001] The present disclosure relates to the technical field of DNA synthesis, and particularly relates to a liquid path supply system and method for DNA synthesis and a DNA synthesis system. BACKGROUND

[0002] Currently, the main technologies for DNA synthesis based on chemical synthesis method on the market are column synthesis method and chip synthesis method. In the column synthesis method, the liquid path design is usually that each reagent passes through an independent pipeline, different reagents are selected by a movement mechanism to be injected into a synthesis column according to the process sequence, and various reactions are performed. The disadvantage of this method is that the higher the throughput is, the more pipelines need to be set, resulting in complex equipment structure, increased cost, and increased maintenance difficulty. In addition, after the throughput is increased, a large amount of reagents flow in the numerous pipelines, and the common valves and connecting pieces cannot completely drain the residual reagents in the pipelines when switching reagents, thereby causing cross contamination between different reagents. The cross contamination seriously affects the quality and accuracy of DNA synthesis, causes errors or impurities in the synthesized DNA fragments, and affects subsequent experiments and applications.

[0003] The liquid path design of the chip synthesis method is more complex than that of the column synthesis method, because all reactions are performed on a surface. It is a difficult problem to switch different reagents without pollution and ensure the cleaning effect. There are usually two methods. One is that the chip surface is open, different reagents are poured onto the chip surface through independent pipelines for reaction, and the reagents on the chip surface are then completely removed by gas purging. This method causes serious pollution due to the open environment. The other is that the reaction is performed in a closed reaction chamber. To avoid cross contamination, the reaction chamber is usually designed with only one inlet, and different reagents are injected through a common pipeline. In the prior art, the liquid supply pipeline is arranged by the column synthesis method, which causes the same problem that the common valves and connecting pieces cannot completely drain the residual reagents in the pipelines when switching reagents, resulting in cross contamination between different reagents and unable to realize the non-pollution switching of different reagents to the common pipeline. SUMMARY

[0004] Therefore, the present disclosure provides a liquid path supply system and method for DNA synthesis and a DNA synthesis system, which can realize the non-pollution switching of different reagents to the corresponding common pipeline of the closed reaction chamber.

[0005] In a first aspect, the present disclosure provides a liquid path supply system for DNA synthesis, comprising:

[0006] a reaction reagent supply system for independent supply of different reaction reagents;

[0007] A switching system comprises a multi-return valve island and a reagent switching assembly installed on the multi-return valve island, the reagent switching assembly being used to control independent switching supply of different reaction reagents and different cleaning reagents;

[0008] The multi-return valve island is provided with a common outlet and a plurality of inlets matched with the reaction reagent supply system and the cleaning source supply system on the side thereof; the multi-return valve island is internally provided with an input branch channel corresponding to each inlet, a delivery branch channel provided corresponding to each input branch channel, and an output main channel communicated with each delivery branch channel, one end of the input branch channel being communicated with the corresponding inlet and the other end being connected with the switching system; the delivery branch channel is connected with the outlet of the switching system;

[0009] The outlets of all the delivery branch channels converge in the same intersection area and the intersection area is communicated with the inlet of the output main channel, and the outlet of the output main channel is communicated with the common outlet;

[0010] The cleaning source supply system is used to supply cleaning reagents and inert gases; the cleaning source supply system comprises a reaction reagent cleaning system matched with the reaction reagent supply system and a valve island cleaning system matched with the switching system.

[0011] In a second aspect, the embodiments of the present disclosure further provide a liquid path supply method for DNA synthesis, based on the liquid path supply system for DNA synthesis, comprising:

[0012] Controlling the reaction reagent supply system and the switching system to supply corresponding reaction reagents;

[0013] Controlling the switching system and the reaction reagent cleaning system to independently clean the reaction reagents flowing through the pipeline and to fill the pipeline with inert gases;

[0014] Controlling the switching system and the valve island cleaning system to clean the internal common pipeline of the multi-return valve island.

[0015] In a third aspect, the embodiments of the present disclosure further provide a DNA synthesis system, comprising a sealed reaction unit, a waste liquid collection unit connected with the outlet of the sealed reaction unit, and a supply system connected with the inlet of the sealed reaction unit through a common pipeline, the supply system being the liquid path supply system for DNA synthesis;

[0016] The supply system is used to supply reaction reagents to the sealed reaction unit, and DNA synthesis is performed in the sealed reaction unit;

[0017] The supply system is used to supply the cleaning reagent after the reaction in the closed reaction unit is completed to clean the pipeline;

[0018] The supply system is used to supply the inert gas after the cleaning of the closed reaction unit is completed to clean the pipeline.

[0019] The liquid supply system for DNA synthesis provided by the embodiments of the present disclosure comprises a reaction reagent supply system, a cleaning source supply system and a switching system, the reaction reagent supply system is used for independent supply of different reaction reagents; the cleaning source supply system is used for supply of cleaning reagents and inert gas; the cleaning source supply system comprises a reaction reagent cleaning system matched with the reaction reagent supply system and a valve island cleaning system matched with the switching system; the switching system comprises a multi-return valve island and a reagent switching assembly installed on the multi-return valve island, the reagent switching assembly is used for controlling independent switching supply of different reaction reagents and different cleaning reagents, the multi-return valve island is provided with a common outlet on the side and a plurality of inlets matched with the reaction reagent supply system and the cleaning source supply system; the multi-return valve island is internally provided with an input branch channel corresponding to each inlet, a conveying branch channel provided corresponding to each input branch channel and an output main channel communicated with each conveying branch channel, one end of the input branch channel is communicated with the corresponding inlet, and the other end is connected with the switching system; the conveying branch channel is connected with the outlet of the switching system; the outlets of all the conveying branch channels converge in the same intersection area, the intersection area is communicated with the inlet of the output main channel, and the outlet of the output main channel is communicated with the common outlet. After each type of reaction reagent is cleaned by the corresponding dedicated reaction reagent cleaning system, the multi-return valve island internal flow channel is cleaned by the valve island cleaning system, that is, when the corresponding pipelines of the three types of reaction reagents are cleaned respectively, although the corresponding solenoid valves located on the outside of the multi-return valve island are switched to the corresponding cleaning source supply, the corresponding reaction reagents are brought into the multi-return valve island, but since the valve island cleaning system does not contact all the reaction reagent supply systems, when the valve island cleaning system is started to clean, there is no reaction reagent, that is, the multi-return valve island inside and the subsequent pipeline can be completely cleaned, different reagents can be switched to the corresponding common pipeline of the closed reaction chamber without pollution, and the reaction quality is ensured.

[0020] The above description is only a summary of the technical solutions of the present disclosure, in order to more clearly understand the technical means of the present disclosure, the contents of the specification can be implemented, and in order to make the above and other purposes, characteristics and advantages of the present disclosure more obvious and easy to understand, the following preferred embodiments are described in detail below, and the accompanying drawings are described as follows. BRIEF DESCRIPTION OF DRAWINGS

[0021] In order to more clearly illustrate the technical solutions of the embodiments of the present disclosure, the drawings needed to be used in the embodiments will be briefly introduced as follows. Obviously, the drawings in the following description only constitute some embodiments of the present disclosure, and for those skilled in the art, other drawings can also be obtained based on these drawings without any creative effort.

[0022] Figure 1 A schematic diagram of a framework of a liquid path supply system for DNA synthesis provided by the embodiments of the present disclosure.

[0023] Figure 2 A schematic diagram of a switching system in Figure 1

[0024] Figure 3 A perspective view of a multi-unification valve island in Figure 2

[0025] Figure 4 Another angle perspective view of a multi-unification valve island in Figure 2

[0026] Figure 5 A schematic diagram of a framework of a DNA synthesis system provided by the embodiments of the present disclosure.

[0027] Explanation of reference signs:

[0028] 10, supply system; 20, closed reaction unit; 30, waste liquid collection unit;

[0029] 100, switching system; 110, multi-unification valve island; 111, input branch passage; 1111, first branch; 1112, second branch; 1113, third branch; 1114, fourth branch; 1115, fifth branch; 1116, sixth branch; 112, delivery branch passage; 113, output main passage; 1141, first inlet; 1142, second inlet; 1143, third inlet; 1144, fourth inlet; 1145, fifth inlet; 1146, sixth inlet; 115, common outlet; 121, first three-way valve; 122, second three-way valve; 123, third three-way valve; 130, inlet joint; 140, outlet joint;

[0030] 210, cap reagent supply unit; 220, oxidizing agent supply unit; 230, deprotection agent supply unit; 240, coupling reagent supply unit;

[0031] 311, first valve island flow passage cleaning unit; 312, second valve island flow passage cleaning unit; 313, third valve island flow passage cleaning unit; 321, cap reagent cleaning unit; 322, oxidizing agent cleaning unit; 323, deprotection agent cleaning unit; 324, coupling reagent cleaning unit. ​​​DETAILED DESCRIPTION

[0032] The embodiments of the present disclosure will be described in detail below with reference to the drawings.

[0033] It should be apparent that the following description illustrates by way of example only a specific embodiment of the present disclosure, and that other advantages and benefits will be apparent to those skilled in the art from this disclosure. It should be noted that the described embodiments are merely a part of the embodiments of the present disclosure, and not all of the embodiments. The present disclosure can also be implemented or applied by other different specific embodiments, and the details in the specification can be modified or changed based on different views and applications without departing from the spirit of the present disclosure. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict. Based on the embodiments in the present disclosure, all other embodiments obtained by those of ordinary skill in the art without creative labor are within the scope of protection of the present disclosure.

[0034] It should be noted that the various aspects of the embodiments described below are within the scope of the appended claims. It should be apparent that the aspects described herein can be embodied in a wide variety of forms and that any specific structure and / or function described herein is merely illustrative. Based on the teachings provided herein one skilled in the art will appreciate that one or more aspects described herein can be implemented independently of any other aspects and that non-dependent aspects can be implemented in conjunction with each other in any way. For example, a device can be implemented using any number of the aspects described herein. Additionally, the methods described herein can be implemented using any number of the aspects described herein.

[0035] It should also be noted that the drawings provided in the following embodiments are only to illustrate the basic concept of the present disclosure, and only the components related to the present disclosure are shown in the drawings, not the number, shape and size of the components when actually implemented. The actual implementation of each component can be a random change in shape, number and proportion, and the layout of the components can also be more complex.

[0036] In addition, in the following description, specific details are provided in order to facilitate a thorough understanding of the examples. However, one skilled in the art will understand that the aspects described can be practiced without these specific details.

[0037] Reference Figure 1 and Figure 2The application discloses a liquid path supply system for DNA synthesis, and relates to the technical field of DNA synthesis, in particular to a liquid path supply system for DNA synthesis.

[0038] The switching system 100 comprises a multi-return valve island 110 and a reagent switching assembly installed on the multi-return valve island 110, and the reagent switching assembly is used for controlling independent switching supply of different reaction reagents and different cleaning reagents; specifically, the reagent switching assembly comprises a plurality of valve control members, and in the embodiment, the plurality of valve control members are preferably a first three-way valve 121, a second three-way valve 122 and a third three-way valve 123.

[0039] The cleaning source supply system is used for supplying cleaning reagents and inert gases, and comprises a reaction reagent cleaning system matched with the reaction reagent supply system and a valve island cleaning system matched with the switching system 100.

[0040] Meanwhile referring to Figure 3 and Figure 4 , the multi-return valve island 110 is provided with a common outlet 115 and a plurality of inlets matched with the reaction reagent supply system and the cleaning source supply system on the side surface, wherein the outlet 115 is provided with an outlet joint 140, and the inlets are respectively provided with inlet joints 130.

[0041] The multi-return valve island 110 is internally provided with an input branch channel 111 corresponding to each inlet, a conveying branch channel 112 arranged corresponding to each input branch channel 111 and an output main channel 113 in communication with each conveying branch channel 112, one end of the input branch channel 111 is in communication with the corresponding inlet, and the other end is connected with the switching system 100; the conveying branch channel 112 is connected with the outlet of the switching system 100.

[0042] The outlets of all the conveying branch channels 112 converge at the same intersection area, and the intersection area is in communication with the inlet of the output main channel 113; the outlet of the output main channel 113 is in communication with the common outlet 115, that is, in the embodiment, the outlets of all the conveying branch channels 112 converge at the inside of the multi-return valve island 110.

[0043] The number of the delivery branch channels 112 is consistent with the number of the valve control members, and each delivery branch channel 112 is connected to the outlet of one valve control member. The lengths of all the delivery branch channels 112 are the same, which can meet the cleaning of the pipelines of the corresponding reaction reagents and simplify the internal structure and facilitate the processing.

[0044] In this embodiment, the reaction reagent cleaning system preferably comprises a deprotection agent cleaning unit 323, an oxidizing agent cleaning unit 322, a capping agent cleaning unit 321, and a coupling reagent cleaning unit 324. The deprotection agent cleaning unit 323 and the deprotection agent supply unit 230 are selectively connected to the inlet of the multi-universal valve island 110 and controlled by an electromagnetic valve. After the deprotection agent supply unit 230 finishes supplying, the deprotection agent cleaning unit 323 can be activated to supply a cleaning reagent (preferably acetonitrile) to clean the pipelines that do not enter the multi-universal valve island 110, the corresponding channels inside the multi-universal valve island 110, and the corresponding areas of the reaction chamber.

[0045] The oxidizing agent cleaning unit 322 and the oxidizing agent supply unit 220 are selectively connected to the inlet of the multi-universal valve island 110 and controlled by an electromagnetic valve. After the oxidizing agent supply unit 220 finishes supplying, the oxidizing agent cleaning unit 322 can be activated to supply a cleaning reagent (preferably acetonitrile) to clean the pipelines that do not enter the multi-universal valve island 110, the corresponding channels inside the multi-universal valve island 110, and the corresponding areas of the reaction chamber.

[0046] The capping agent cleaning unit 321 and the capping agent supply unit 210 are selectively connected to the inlet of the multi-universal valve island 110 and controlled by an electromagnetic valve. After the capping agent supply unit 210 finishes supplying, the capping agent cleaning unit 321 can be activated to supply a cleaning reagent (preferably acetonitrile) to clean the pipelines that do not enter the multi-universal valve island 110, the corresponding channels inside the multi-universal valve island 110, and the corresponding areas of the reaction chamber.

[0047] The coupling reagent cleaning unit 324, the coupling reagent supply unit 240 and the inlet of the multi-uni valve island 110 are in alternative communication and controlled by electromagnetic valves. After the coupling reagent supply unit 240 finishes supplying, the coupling reagent cleaning unit 324 can be activated to supply cleaning reagent (preferably acetonitrile) to clean the pipes that do not enter the multi-uni valve island 110, the corresponding channels inside the multi-uni valve island 110 and the corresponding areas of the reaction chamber.

[0048] The valve island cleaning system includes a plurality of valve island flow channel cleaning units. The plurality of valve island flow channel cleaning units are respectively arranged corresponding to the plurality of valve control members and do not interfere with the reaction reagent supply system. In this embodiment, the plurality of valve island flow channel cleaning units are preferably respectively a valve island flow channel first cleaning unit 311, a valve island flow channel second cleaning unit 312 and a valve island flow channel third cleaning unit 313, which are respectively corresponding to the first three-way valve 121, the second three-way valve 122 and the third three-way valve 123, i.e. through the inlet of the first three-way valve 121, the second three-way valve 122 and the third three-way valve 123 to clean the internal flow channel of the multi-uni valve island 110, realizing thorough cleaning of all common pipes.

[0049] In actual operation, due to the defects of the structure of the electromagnetic valve itself, when the corresponding cleaning unit is opened, the corresponding reaction reagent will be brought out, leading to the entry into the common pipe, which cannot realize thorough cleaning of the common pipe, and further leading to cross contamination of different reagents. Through the setting of the cleaning unit corresponding to each type of reaction reagent in this application and the three independent valve island flow channel cleaning units, thorough cleaning of the common pipe can be realized, effectively solving the problem of cross contamination of reaction reagents. Specifically, in the system disclosed in this application, after cleaning by the exclusive reaction reagent cleaning system corresponding to each type of reaction reagent, the internal flow channel of the multi-uni valve island 110 is cleaned by the valve island cleaning system, i.e. when the corresponding pipes of the three types of reaction reagents are cleaned respectively, although the corresponding electromagnetic valve located outside the multi-uni valve island 110 is switched to the corresponding cleaning source supply, the corresponding reaction reagent will be brought into the multi-uni valve island 110, but since the valve island cleaning system does not contact all the reaction reagent supply systems, when the valve island cleaning system is activated for cleaning, no reaction reagent will appear, i.e. thorough cleaning of the multi-uni valve island 110 and the subsequent pipes can be realized.

[0050] In this embodiment, the angles between the two adjacent conveying branch channels 112 are acute angles, so that the conveying branch channels 112 are elongated channels inside the multi-return-to-one valve island 110, and it is ensured that the various reagents inside the multi-return-to-one valve island 110 mostly flow in the corresponding channels, and when flowing out of the corresponding conveying branch channels 112, the reagents quickly flow out of the output main channel 113 without flowing back to other conveying branch channels 112 or the corresponding three-way valves of other conveying branch channels 112, so as not to contaminate the three-way valves provided on the multi-return-to-one valve island 110.

[0051] In this embodiment, the several entrances provided on the side of the multi-return-to-one valve island 110 are respectively a first group of entrances, a second group of entrances, and a third group of entrances. The first group of entrances includes oppositely arranged first and second entrances 1141 and 1142. The first entrance 1141 is in selective communication with the cap reagent cleaning unit 321 and the cap reagent supply unit 210. The second entrance 1142 is in selective communication with the coupling reagent cleaning unit 324, the coupling reagent supply unit 240, and the first cleaning unit.

[0052] The second group of entrances includes oppositely arranged third and fourth entrances 1143 and 1144. The third entrance 1143 is in selective communication with the oxidizing agent cleaning unit 322 and the oxidizing agent supply unit 220. The fourth entrance 1144 is connected to the second cleaning unit.

[0053] The third group of entrances includes oppositely arranged fifth and sixth entrances 1145 and 1146. The fifth entrance 1145 is in selective communication with the deprotection agent cleaning unit 323 and the deprotection agent supply unit 230. The sixth entrance 1146 is connected to the third cleaning unit.

[0054] In this embodiment, the cap reagent supply unit 210 and the coupling reagent supply unit 240 correspond to the first three-way valve 121 in the multi-return-to-one valve island 110. Even if there is residual coupling reagent in the first three-way valve 121, it will be covered by the cap reagent, so it will not have an impact. Only the cap reagent has this effect on the coupling reagent, and other reaction reagents will affect each other. That is, only these two types of reaction reagents can share an electromagnetic valve, and other reaction reagents each need to correspond to an independent electromagnetic valve.

[0055] Further, the first, third, and fifth entrances 1141, 1143, and 1145 are provided on the same side of the multi-return-to-one valve island 110. The normally open entrances of the first, second, and third three-way valves 121, 122, and 123 are respectively in communication with the first, second, and third cleaning units, which is convenient for control.

[0056] In this embodiment, the three delivery branch channels 112 are provided, and the three delivery branch channels 112 are respectively communicated with the outlets of the first three-way valve 121, the second three-way valve 122 and the third three-way valve 123; the three delivery branch channels 112 are straight section holes, and the included angle between adjacent two delivery branch channels 112 is not greater than 30°, the lengths of the three delivery branch channels 112 are the same, and are greater than the length of the output main channel 113, that is, it is ensured that the reaction reagent in each channel will not be shunted to the internal part of the electromagnetic valve in other channels.

[0057] The input branch channel 111 is an L-shaped hole, and the L-shaped hole includes a first branch section communicated with an inlet and a second branch section connected with a valve control member; the input branch channel 111 provided in the multi-return valve island 110 includes first, second, third, fourth, fifth and sixth branch channels 1111, 1112, 1113, 1114, 1115 and 1116 respectively communicated with first, second, third, fourth, fifth and sixth inlets 1141, 1142, 1143, 1144, 1145 and 1146; the second branch section in the first branch channel 1111 is arranged in parallel and equal length with the second branch section in the second branch channel 1112; the second branch section in the third branch channel 1113 is arranged in parallel and equal length with the second branch section in the fourth branch channel 1114; the second branch section in the fifth branch channel 1115 is arranged in parallel and equal length with the second branch section in the sixth branch channel 1116, which not only meets the assembly requirement, but also ensures the tight installation with the corresponding electromagnetic valve.

[0058] The three delivery branch channels 112 are respectively arranged in parallel with the second branch section in the first branch channel 1111, the second branch section in the third branch channel 1113 and the second branch section in the fifth branch channel 1115, and the length of the delivery branch channel 112 is greater than the length of the second branch section, that is, it is ensured that the common pipeline of different reagents in the multi-return valve island 110 is relatively short, unnecessary pipelines are reduced, and the reaction cavity inside is supplied as soon as possible.

[0059] The deprotection agent cleaning unit 323, the oxidizing agent cleaning unit 322, the cap agent cleaning unit 321, the coupling reagent cleaning unit 324, the first cleaning unit, the second cleaning unit and the third cleaning unit all include a cleaning reagent supply source and an inert gas supply source; the deprotection agent cleaning unit 323 is controlled by the first multi-way valve through the deprotection agent supply unit 230, the oxidizing agent cleaning unit 322 is controlled by the second multi-way valve through the oxidizing agent supply unit 220, the cap agent cleaning unit 321 is controlled by the third multi-way valve through the cap agent supply unit 210, and the coupling reagent cleaning unit 324 is controlled by the fourth multi-way valve through the coupling reagent supply unit 240.

[0060] In this embodiment, inert gas is filled in all pipelines before cleaning reagent cleaning, so that when cleaning the pipeline corresponding to a certain type of reaction reagent, the inert gas filled in other pipelines can also ensure that the solenoid valve in other channels will not backflow into the multi-return valve island 110, ensuring thorough cleaning of the multi-return valve island 110 and subsequent pipelines.

[0061] Referring to Figs. 1 and 2 simultaneously, Figure 5 The application also discloses a DNA synthesis system, which comprises a closed reaction unit 20, a waste liquid collection unit 30 connected with the outlet of the closed reaction unit 20, a supply system 10 connected with the inlet of the closed reaction unit 20 through a common pipeline, the supply system 10 being used for supplying reaction reagents to the closed reaction unit 20 to perform DNA synthesis in the closed reaction unit 20, the supply system 10 being used for supplying cleaning reagents to the closed reaction unit 20 to clean the pipeline after the reaction in the closed reaction unit 20, and the supply system 10 being used for supplying inert gas to the closed reaction unit 20 to purge the pipeline after the cleaning of the closed reaction unit 20.

[0062] In this embodiment, the deprotection agent cleaning unit, the oxidizing agent cleaning unit and the capping agent cleaning unit share a cleaning reagent supply source and an inert gas supply source, the cleaning reagent supply source is preferably acetonitrile, and the inert gas supply source is preferably nitrogen, the acetonitrile and the nitrogen are connected with two inlets of a three-way valve respectively and then connected with an inlet of a four-way valve, the outlet ends of the deprotection agent supply unit, the oxidizing agent supply unit and the capping agent supply unit are respectively connected with a two-way valve and a three-way valve, the corresponding reaction reagent can be controlled to be supplied through the corresponding two-way valve, the three three-way valves on the supply pipelines of the three types of reaction reagents are respectively connected with three outlets of the four-way valve, and the cleaning source supply of the corresponding pipeline can be controlled through the four-way valve to perform independent cleaning of the corresponding pipeline. Finally, the outlets of the three three-way valves on the supply pipelines of the three types of reaction reagents are respectively connected with corresponding inlets of a first three-way valve, a second three-way valve and a third three-way valve.

[0063] The acetonitrile and the nitrogen are switched through the corresponding three-way valves, the corresponding three-way valves are always open, the nitrogen pipeline is provided with a nitrogen cut-off valve, and the cut-off valve is opened when the nitrogen is supplied.

[0064] The capping agent cleaning unit comprises a capping agent A supply source, a capping agent B supply source and a mixing three-way valve, the capping agent A and the capping agent B are respectively supplied through the capping agent A supply source and the capping agent B supply source, the two are mixed in a 1:1 ratio and used, the capping agent is formed through the mixing three-way valve, and then connected to the corresponding inlet of the first three-way valve through a pipeline.

[0065] In the embodiment, the several valve island flow channel cleaning units also share the cleaning reagent supply source and the inert gas supply source. The cleaning reagent supply source is preferably acetonitrile, and the inert gas supply source is preferably nitrogen. The acetonitrile and the nitrogen are connected to two inlets of a three-way valve, and then connected to an inlet of a four-way valve. Three outlets of the four-way valve are connected to corresponding inlets of the first three-way valve, the second three-way valve and the third three-way valve through corresponding pipelines.

[0066] The coupling reagent supply unit includes a T monomer supply source, an activator supply source and a mixing three-way valve. The T monomer and the activator are supplied through the T monomer supply source and the activator supply source respectively, mixed in a 1:1 ratio, and then formed into a coupling reagent through the mixing three-way valve. The coupling reagent is then connected to a corresponding inlet of the first three-way valve through a pipeline.

[0067] The application also discloses a liquid path supply method for DNA synthesis, based on the liquid path supply system for DNA synthesis.

[0068] S100, controlling the reaction reagent supply system and the switching system to supply corresponding reaction reagents;

[0069] S200, controlling the switching system and the reaction reagent cleaning system to independently clean and inert gas purge and fill the corresponding reaction reagent flow through the pipeline.

[0070] S300, controlling the switching system and the valve island cleaning system to clean the internal common pipeline of the multi-return valve island.

[0071] In the application, by setting the special reaction reagent cleaning system corresponding to each type of reaction reagent and the three independent valve island flow channel cleaning units, when cleaning the corresponding pipeline of each type of reaction reagent, even if the corresponding electromagnetic valve outside the multi-return valve island is switched to the cleaning source supply and the reaction reagent is brought into the multi-return valve island, the subsequent cleaning is carried out through the valve island cleaning system. Since the valve island cleaning system does not contact all the reaction reagent supply systems, the reaction reagent is not brought out, so that the multi-return valve island and the subsequent pipeline are thoroughly cleaned, cross contamination of different reagents is avoided, and the quality and accuracy of DNA synthesis are improved.

[0072] By controlling the independent switching and supply of different reaction reagents and different cleaning reagents through the reagent switching assembly, and combining the unique flow channel design of the multi-return valve island, residual reagents can be more effectively avoided from entering the common pipeline when switching reagents, different reagents are ensured to be switched to the common pipeline without pollution, DNA fragment errors or impurity problems caused by cross contamination are reduced, and reliable DNA samples are provided for subsequent experiments and applications.

[0073] Unlike the situation that the higher the flux is, the more the pipelines are in the traditional column synthesis method, the application adopts a multi-universal valve island design to integrate and supply different reaction reagents and cleaning reagents through the multi-universal valve island 110. Although there are complex flow channels inside the multi-universal valve island, the use of the number of external independent pipelines is effectively reduced as a whole, the liquid path structure of the equipment is simplified, the difficulty of equipment design and manufacturing is reduced, the cost of raw materials is reduced, and the probability of failure is effectively reduced due to fewer pipelines and more reasonable flow channel layout. When troubleshooting and maintaining the failure, it is easier to locate the problem, and the time and effort required for maintenance are reduced. At the same time, due to the effective avoidance of cross-contamination, the damage of the equipment and the waste of reagents caused by pollution are reduced, and the maintenance cost is further reduced.

[0074] The application can provide a more reliable liquid path supply scheme for chip synthesis method. For the problems of open environment easy to be contaminated and cross-contamination of public pipelines in the closed reaction cavity in the chip synthesis method, the application can better meet the requirements of the chip synthesis method for different reagents without contamination switching and cleaning effect through effective cleaning and switching design, and has wider applicability.

[0075] The detailed description of the present embodiment can refer to the corresponding description in the foregoing embodiments, which will not be repeated here.

[0076] The basic principles of the present disclosure are described above in combination with specific embodiments, but it should be pointed out that the advantages, advantages, effects and the like mentioned in the present disclosure are only examples and not limitations, and these advantages, advantages, effects and the like cannot be considered as the must-have of each embodiment of the present disclosure. In addition, the specific details of the above disclosure are only for the purpose of example and for the purpose of understanding, and the above details do not limit the present disclosure to the must-use specific details.

[0077] In the present disclosure, the relationship terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between the entities or operations. The block diagrams of devices, apparatuses, equipment, systems involved in the present disclosure are only illustrative examples and are not intended to require or imply the connection, arrangement, configuration shown in the block diagram. As those skilled in the art will recognize, these devices, apparatuses, equipment, systems can be connected, arranged, configured in any manner. Words such as "include", "contain", "have" and the like are open-ended words, which mean "include but not limited to", and can be used interchangeably. The words "or" and "and" used herein mean the word "and / or", and can be used interchangeably unless the context clearly indicates otherwise. The word "such as" used herein means the phrase "such as but not limited to", and can be used interchangeably.

[0078] Also, as used in the description herein and throughout the claims that follow, the meaning of "or" includes "and / or" unless the context clearly dictates otherwise. For example, when used in the examination of "at least one of A or / and B" the phrase "at least one of A or B" means A or B or both. As used in the examination of "between one and three of A, B, and C," the phrase means A or B or C, or any combination thereof.

[0079] It is also important to note that the systems and methods of the disclosure can be embodied in a variety of forms depending on the particular requirements of the applications. Some embodiments can be implemented in software, while other embodiments can be implemented in hardware or a combination of software and hardware. As used herein the term "software" includes but is not limited to one or more computer program(s) or other computer readable instructions that can be stored or carried on a computer readable medium, such as a floppy disk, a flexible disk, a hard disk, a magnetic tape, a magnetic disk, an optical disk, a CD-ROM, RAM, ROM, PROM, EPROM, a flash memory, a solid state drive, or other memory chip or cartridge, or the like.

[0080] Various changes, modifications and alterations to the techniques described herein can be made without departing from the teachings of the attached claims. Moreover, the scope of the claims should not be limited to the particular aspects described in the above description and drawings. Rather, the scope of the claims should be understood to include all aspects and equivalents thereof that are within the scope of the claims.

[0081] The above description of disclosed aspects is intended to be illustrative, and not restrictive. Many other aspects can be apparent to those of skill in the art upon reviewing the above description. The scope of the claims should, therefore, be interpreted to include all such aspects and their equivalents.

[0082] The above description has been given by way of example and is not intended to limit the application. Although various example aspects and embodiments have been discussed, those skilled in the art will recognize that certain modifications, substitutions, changes, additions and omissions can be made without departing from the scope of the application.

Claims

1. A fluidic supply system for DNA synthesis, characterized by, The application relates to a reaction reagent supply system, a cleaning source supply system, a switching system and a multi-return valve island. The reaction reagent supply system is used for independent supply of different reaction reagents. The cleaning source supply system is used for supply of cleaning reagents and inert gases. The switching system comprises a multi-return valve island and a reagent switching assembly arranged on the multi-return valve island, and the reagent switching assembly is used for controlling independent switching supply of different reaction reagents and different cleaning reagents. The multi-return valve island is provided with a common outlet and a plurality of entrances matched with the reaction reagent supply system and the cleaning source supply system. The multi-return valve island is internally provided with input branch channels corresponding to the entrances respectively, conveying branch channels arranged corresponding to the input branch channels and an output main channel communicated with the conveying branch channels. One end of the input branch channel is communicated with the corresponding entrance, and the other end is connected with the switching system. The conveying branch channels are connected with the outlet of the switching system. The outlets of all the conveying branch channels converge in a same convergence area, and the convergence area is communicated with the inlet of the output main channel. The outlet of the output main channel is communicated with the common outlet. The cleaning source supply system comprises a reaction reagent cleaning system matched with the reaction reagent supply system and a valve island cleaning system matched with the switching system. The plurality of entrances of the multi-return valve island are respectively a first group of entrances, a second group of entrances and a third group of entrances.

2. The liquid path supply system for DNA synthesis according to claim 1, wherein The first group of entrances comprises oppositely arranged first and second entrances. The second group of entrances comprises oppositely arranged third and fourth entrances.

3. The liquid path supply system for DNA synthesis according to claim 2, wherein The third group of entrances comprises oppositely arranged fifth and sixth entrances.

4. The liquid path supply system for DNA synthesis according to claim 2, wherein The input branch channel is an L-shaped channel, which comprises a first branch section communicated with the entrance and a second branch section connected with the valve control member. The input branch channel internally arranged in the multi-return valve island comprises first, second, third, fourth, fifth and sixth branch channels communicated with the first, second, third, fourth, fifth and sixth entrances respectively. The second branch section in the first branch channel is arranged in parallel with the second branch section in the second branch channel. The second branch section in the third branch channel is arranged in parallel with the second branch section in the fourth branch channel. The second branch section in the fifth branch channel is arranged in parallel with the second branch section in the sixth branch channel. The lengths of the conveying branch channels are greater than the lengths of the second branch sections. The number of the conveying branch channels is consistent with the number of the valve control members, and each conveying branch channel is communicated with the outlet of one valve control member. The lengths of all the conveying branch channels are the same. The included angles between adjacent two conveying branch channels are acute angles. The reaction reagent supply system comprises a deprotection agent supply unit, an oxidizing agent supply unit, a cap agent supply unit and a coupling reagent supply unit. The reaction reagent cleaning system comprises a deprotection agent cleaning unit, an oxidizing agent cleaning unit, a capping agent cleaning unit, and a coupling reagent cleaning unit; the deprotection agent cleaning unit and the deprotection agent supply unit are in selective communication with the inlet of the multi-universal valve island; the oxidizing agent cleaning unit and the oxidizing agent supply unit are in selective communication with the inlet of the multi-universal valve island; the capping agent cleaning unit and the capping agent supply unit are in selective communication with the inlet of the multi-universal valve island; and the coupling reagent cleaning unit and the coupling reagent supply unit are in selective communication with the inlet of the multi-universal valve island. The valve island cleaning system comprises a plurality of valve island flow channel cleaning units; the plurality of valve island flow channel cleaning units are respectively arranged in correspondence with the plurality of valve controls and do not interfere with the reaction reagent supply system.

5. The liquid path supply system for DNA synthesis according to claim 4, wherein The plurality of valve controls comprise a first three-way valve, a second three-way valve, and a third three-way valve. The plurality of valve island flow channel cleaning units comprise a first cleaning unit, a second cleaning unit, and a third cleaning unit which are independently arranged and respectively matched with the first three-way valve, the second three-way valve, and the third three-way valve. The first inlet is in selective communication with the capping agent cleaning unit and the capping agent supply unit; the second inlet is in selective communication with the coupling reagent cleaning unit, the coupling reagent supply unit, and the first cleaning unit; The third inlet is in selective communication with the oxidizing agent cleaning unit and the oxidizing agent supply unit; the fourth inlet is connected with the second cleaning unit; The fifth inlet is in selective communication with the deprotection agent cleaning unit and the deprotection agent supply unit; and the sixth inlet is connected with the third cleaning unit. The three delivery branch channels are respectively in communication with the outlets of the first three-way valve, the second three-way valve, and the third three-way valve.

6. The liquid path supply system for DNA synthesis according to claim 5, wherein The first inlet, the third inlet, and the fifth inlet are arranged on the same side of the multi-universal valve island. The normally open inlets of the first three-way valve, the second three-way valve, and the third three-way valve are respectively in communication with the first cleaning unit, the second cleaning unit, and the third cleaning unit.

7. The liquid path supply system for DNA synthesis according to claim 5, wherein The deprotection agent cleaning unit, the oxidizing agent cleaning unit, the capping agent cleaning unit, the coupling reagent cleaning unit, the first cleaning unit, the second cleaning unit, and the third cleaning unit all comprise a cleaning reagent supply source and an inert gas supply source. The deprotection agent cleaning unit and the deprotection agent supply unit are controlled by a first multi-way valve; The oxidizing agent cleaning unit and the oxidizing agent supply unit are controlled by a second multi-way valve; The capping agent cleaning unit and the capping agent supply unit are controlled by a third multi-way valve; The coupling reagent cleaning unit and the coupling reagent supply unit are controlled by a fourth multi-way valve.

8. A liquid path supply method for DNA synthesis, characterized by, The liquid path supply system for DNA synthesis according to any one of claims 1-7, comprising: controlling the reaction reagent supply system and the switching system to supply corresponding reaction reagents; controlling the switching system and the reaction reagent cleaning system to independently clean and inert gas purge fill the corresponding reaction reagents flowing through the pipeline; The switching system and the valve island cleaning system are controlled to clean the internal common pipeline of the multi-return valve island.

9. A DNA synthesis system, characterized by, The system comprises a closed reaction unit, a waste liquid collection unit connected with the outlet of the closed reaction unit, and a supply system connected with the inlet of the closed reaction unit through a common pipeline, wherein the supply system is any one of the liquid path supply systems for DNA synthesis described in items 1-7. The supply system is used to supply reaction reagents to the closed reaction unit, and DNA synthesis is performed in the closed reaction unit. The supply system is used to supply cleaning reagents after the reaction in the closed reaction unit is completed to clean the pipeline. The supply system is used to supply inert gas after the cleaning of the closed reaction unit is completed to clean the pipeline.

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