Multi-chamber microfluidic chip and DNA synthesis method based on multi-chamber microfluidic chip

Abstract

The invention provides a multi-chamber microfluidic chip and a DNA synthesis method based on the multi-chamber microfluidic chip, the multi-chamber microfluidic chip comprises a microfluidic chip, the microfluidic chip is provided with a synthesis chamber, a uniform mixing chamber, an assembly chamber, a liquid inlet hole, a liquid outlet hole, a micro valve, a microelectrode and a uniform mixer, the synthesis chamber is communicated with the uniform mixing chamber through a micro channel, and the uniform mixing chamber is communicated with the assembly chamber through a micro channel; the blending chamber is communicated with the assembling chamber through a micro-channel, one end, deviating from the blending chamber, of the assembling chamber is communicated with a liquid discharging channel, micro valves are arranged in the micro-channel and the liquid discharging channel, the liquid inlet hole is communicated with the synthesis chamber, the liquid outlet hole is communicated with the liquid discharging channel, a micro-electrode is arranged in the synthesis chamber, and a blending device is arranged in the blending chamber; and the temperature control device is used for regulating and controlling the temperature of the microfluidic chip in the placing groove. In this way, transfer between different devices is not needed in the synthesis process, and therefore alignment errors caused by chip movement are avoided.

Description

Technical Field

[0001] The present invention relates to the technical field of biochips, and in particular to a multi-chamber microfluidic chip and a DNA synthesis method based on the multi-chamber microfluidic chip. Background Art

[0002] In the relevant DNA chip synthesis technology, the synthesis process usually involves multiple steps, such as: synthesis of oligonucleotide arrays, screening and elution, and assembly of long-chain DNA. These steps often require the transfer of microfluidic chips between different devices to complete the reactions at each stage. When the microfluidic chip moves between different devices, it is easy to produce inaccurate alignment, resulting in misalignment of the working site. This misalignment not only reduces the accuracy and efficiency of the synthesis process, but may also affect the quality and yield of the final DNA product. In addition, the relevant DNA synthesis technology often requires a lot of manual participation, including chip transfer, setting, monitoring and adjustment of reaction conditions. This highly manual operation method not only increases the complexity and time cost of the operation, but also significantly increases the error rate caused by human factors. Summary of the invention

[0003] In view of this, the purpose of the present invention is to propose a multi-chamber microfluidic chip and a DNA synthesis method based on the multi-chamber microfluidic chip, which are used to solve the problem of transferring the microfluidic chip between different devices when synthesizing DNA, so as to at least partially solve the problems in the related art.

[0004] Based on the above purpose, a first aspect of an embodiment of the present invention provides a multi-chamber microfluidic chip, comprising:

[0005] A microfluidic chip, comprising a synthesis chamber, a mixing chamber, an assembly chamber, a liquid inlet hole, a liquid outlet hole, a microvalve, a microelectrode and a mixer, wherein the synthesis chamber is connected to the mixing chamber via a microchannel, the mixing chamber is connected to the assembly chamber via the microchannel, an end of the assembly chamber away from the mixing chamber is connected to a drainage channel, the microchannel and the drainage channel are both provided with the microvalve, the liquid inlet hole is connected to the synthesis chamber, the liquid outlet hole is connected to the drainage channel, the microelectrode is provided in the synthesis chamber, and the mixer is provided in the mixing chamber;

[0006] The temperature control device comprises a placement groove for placing the microfluidic chip and a temperature control mechanism, wherein the temperature control mechanism is used to control the temperature of the microfluidic chip located in the placement groove.

[0007] Optionally, the microfluidic chip comprises a sealing plate, a reaction plate, a driving plate and a bottom plate which are fixedly connected in sequence;

[0008] The sealing plate is provided with the liquid inlet and the liquid outlet, the reaction plate is provided with a synthesis through hole, a mixing through hole and an assembly through hole, and the synthesis through hole is connected with the mixing through hole through a flow channel, the mixing through hole and the assembly through hole are connected with the flow channel, and the assembly through hole is provided with a discharge channel at one end away from the flow channel, the driving plate is provided with a synthesis groove, a mixing groove and an assembly groove, the synthesis groove is provided with the microelectrode, the mixing groove is provided with the mixer, the synthesis groove is connected with the mixing groove through a through groove, the mixing groove is connected with the assembly groove through the through groove, and the assembly groove is away from the through groove. A discharge groove is provided at one end of the groove. After the sealing plate, the reaction plate and the driving plate are fixedly connected, the sealing plate, the synthesis through hole and the synthesis groove form the synthesis chamber, the sealing plate, the mixing through hole and the mixing groove form the mixing chamber, the sealing plate, the assembly through hole and the assembly groove form the assembly chamber, the sealing plate, the flow channel and the through groove form the microchannel, the sealing plate, the discharge channel and the discharge groove form the drainage channel, the bottom plate is fixedly connected to the lower surface of the driving plate and a signal control circuit for controlling the microvalve, the microelectrode and the mixer is provided inside the bottom plate.

[0009] Optionally, a heating electrode is provided in the assembly chamber, and the number of the heating electrodes is multiple, and the heating electrode is connected to the signal control circuit.

[0010] Optionally, a plurality of microfluidic walls are arranged at intervals in the synthesis chamber, a microfluidic channel for liquid circulation is formed between two adjacent microfluidic walls, and the microelectrode is provided on the bottom surface of the microfluidic channel.

[0011] Optionally, the mixer comprises a piezoelectric ceramic sheet.

[0012] Optionally, the temperature control device includes a base plate, a cover plate and a fixing assembly;

[0013] The substrate is provided with the placement groove, the cover plate is arranged in the placement groove and placed above the microfluidic chip, and the fixing component is arranged on the upper surface of the substrate and has an abutment member for abutting the cover plate;

[0014] The temperature control mechanism comprises a heat dissipation component and a heating plate. The heat dissipation component is arranged on the lower surface of the substrate. An embedding groove for accommodating the heating plate is arranged on the bottom surface of the placement groove.

[0015] Optionally, the fixing assembly includes a column, a sleeve, a connecting rod and a screw;

[0016] One end of the column is fixedly connected to the upper surface of the base plate, the sleeve is sleeved on the column and the connecting rod is fixedly connected to the side wall of the sleeve, the abutment includes the screw, and the end of the connecting rod away from the sleeve is provided with a threaded hole for threaded connection with the screw.

[0017] Optionally, the heat dissipation assembly includes a heat dissipation plate, the heat dissipation plate is installed on the lower surface of the substrate, there are multiple heat dissipation plates, and the multiple heat dissipation plates are arranged at intervals along the length direction of the substrate;

[0018] It also includes a support frame, a mounting frame and a fan. The edge of the lower surface of the substrate is fixedly connected to a support column for connecting to the mounting frame. The mounting frame is provided with a plug hole for plugging the support column. The fan is installed on the mounting frame and the air outlet of the fan is arranged toward the heat sink. The support frame is placed on the lower surface of the mounting frame, and a rubber gasket is provided on the bottom surface of the support frame.

[0019] Optionally, both ends of the embedding groove that are oppositely arranged along the length direction of the substrate are provided with hand-gripping grooves, the hand-gripping grooves are communicated with the embedding groove, and a through groove for passing a power cord of the heating plate is provided on the substrate, the through groove is communicated with the embedding groove;

[0020] A handle is fixedly connected to the rod head of the screw rod;

[0021] There are multiple fixing components, and the multiple fixing components are arranged at intervals along the edge of the substrate.

[0022] According to a second aspect of an embodiment of the present invention, a method for synthesizing DNA based on a multi-chamber microfluidic chip is provided, wherein DNA is synthesized by the multi-chamber microfluidic chip described in any one of the above embodiments, and the DNA synthesis method comprises:

[0023] Injecting a synthesis reagent into the synthesis chamber through the liquid inlet hole, so that the synthesis reagent undergoes a synthesis reaction with the energized microelectrode, thereby obtaining a target oligonucleotide array;

[0024] Injecting a cleaning agent into the synthesis chamber through the liquid inlet hole, and discharging the cleaning agent after cleaning through the liquid outlet hole;

[0025] Injecting a lysing agent into the synthesis chamber through the liquid inlet hole, electrifying the microelectrode to generate a voltage according to the sequence design, generating an electrochemical reaction to break the chemical bond between the oligonucleotide array and the microelectrode, and lysing the oligonucleotide array;

[0026] Injecting the assembly reagent into the synthesis chamber through the liquid inlet, so that the assembly reagent and the mixed solution containing the oligonucleotide flow into the mixing chamber through the microchannel between the synthesis chamber and the mixing chamber;

[0027] Mixing the assembly reagent and the mixed solution containing the oligonucleotide by the mixer;

[0028] The mixed assembly reagent and the mixed solution containing the oligonucleotide are discharged into the assembly chamber through the microchannel located between the mixing chamber and the assembly chamber to assemble the long-chain DNA, and the temperature is controlled by the temperature control mechanism and / or the heating electrode;

[0029] After the assembly is completed, the mixed solution containing the long-chain DNA is discharged from the assembly chamber through the liquid outlet.

[0030] Through the above technical scheme, the synthesis chamber, mixing chamber and assembly chamber provided inside the microfluidic chip can provide support for the synthesis of long-chain DNA without the need for transfer between different devices, thereby avoiding the alignment error caused by chip movement, reducing manual participation, reducing the mismatch rate and improving the overall efficiency and success rate of DNA synthesis. That is, a microelectrode is provided in the synthesis chamber, and the reagent for synthesizing oligonucleotides is injected into the synthesis chamber through the liquid inlet hole, and then directly contacts the microelectrode in the synthesis chamber. The microelectrode is started by controlling the signal control circuit. After multiple rounds of synthesis reactions, the desired oligonucleotide array is obtained, and then the cleavage agent is injected into the synthesis chamber, and the microelectrode is selected according to the sequence design. Power is turned on to generate voltage, produce electrochemical reaction, and then break the chemical bond between the oligonucleotide and the electrode, cleave the oligonucleotide, and then inject the assembly reagent into the synthesis chamber through the liquid inlet hole, and push the mixture containing the cleaved oligonucleotide in the synthesis chamber into the mixing chamber, and mix the cleaved oligonucleotide mixture and the assembly reagent by the mixer arranged in the mixing chamber so that the two can be evenly mixed. Finally, the evenly mixed mixture is injected into the assembly chamber through the microchannel, and the microfluidic chip is heated by the temperature control device to provide a good environmental temperature for the assembly of long-chain DNA and improve the assembly efficiency. After the assembly is completed, the mixed solution containing the long-chain DNA is discharged from the liquid outlet hole.

[0031] Other features and advantages of the present disclosure will be described in detail in the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0033] Figure 1 is a schematic structural diagram of a microfluidic chip and a temperature control device provided in an exemplary embodiment of the present disclosure;

[0034] Figure 2 is a schematic structural diagram of a microfluidic chip provided in an exemplary embodiment of the present disclosure;

[0035] Figure 3 is a schematic structural diagram of a temperature control device provided in an exemplary embodiment of the present disclosure;

[0036] Figure 4 is a schematic structural diagram of a mounting frame and a fan provided in an exemplary embodiment of the present disclosure;

[0037] Figure 5 is a schematic structural diagram of a sealing plate provided in an exemplary embodiment of the present disclosure;

[0038] Figure 6 is a schematic structural diagram of a reaction plate provided in an exemplary embodiment of the present disclosure;

[0039] Figure 7 is a schematic structural diagram of a driving board provided in an exemplary embodiment of the present disclosure;

[0040] Figure 8 is a schematic structural diagram of a base plate provided in an exemplary embodiment of the present disclosure;

[0041] Fig. 9 Schematic diagram of a process for DNA synthesis based on a multi-chamber microfluidic chip provided in an exemplary embodiment of the present disclosure.

[0042] Description of Reference Numerals

[0043] 1-microfluidic chip; 101-sealing plate; 1011-liquid inlet hole; 1012-liquid outlet hole; 102-reaction plate; 1021-synthetic through hole; 1022-mixing through hole; 1023-assembly through hole; 1024-flow channel; 1025-discharge channel; 103-driving plate; 1031-synthetic tank; 1032-mixing tank; 1033-assembly tank; 1034-through tank; 1035-discharge tank; 104-bottom plate; 105-microvalve; 106-microelectrode; 107-mixer; 2-temperature control device; 201- Temperature control mechanism; 2011-heating plate; 2012-heating plate; 202-base plate; 2021-placing slot; 2022-embedding slot; 2023-handle slot; 2024-through slot; 203-cover plate; 204-fixing assembly; 2041-column; 2042-sleeve; 2043-connecting rod; 2044-screw; 3-heating electrode; 4-signal control circuit; 5-microfluidic wall; 501-microchannel; 6-support frame; 7-mounting frame; 701-fan; 702-plug hole; 703-support column; 8-handle. DETAILED DESCRIPTION

[0044] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific embodiments and with reference to the accompanying drawings.

[0045] It should be noted that, unless otherwise defined, the technical terms or scientific terms used in the embodiments of the present invention should be understood by people with ordinary skills in the field to which the present disclosure belongs. The "first", "second" and similar words used in the present disclosure do not represent any order, quantity or importance, but are only used to distinguish different components. "Including" or "comprising" and similar words mean that the elements or objects appearing in front of 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 can 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.

[0046] In the specific embodiment provided in the present disclosure, a multi-chamber microfluidic chip 1 is provided, referring to Figures 1 to 8As shown, the multi-chamber microfluidic chip 1 comprises: a microfluidic chip 1 and a temperature control device 2, wherein the microfluidic chip 1 has a synthesis chamber, a mixing chamber, an assembly chamber, a liquid inlet 1011, a liquid outlet 1012, a microvalve 105, a microelectrode 106 and a mixer 107, the synthesis chamber is connected to the mixing chamber through a microchannel, the mixing chamber is connected to the assembly chamber through a microchannel, the end of the assembly chamber away from the mixing chamber is connected to a drainage channel, the microchannel and the drainage channel are both provided with microvalves 105, the liquid inlet 1011 is connected to the synthesis chamber, The liquid outlet 1012 is connected to the drainage channel, a microelectrode 106 is provided in the synthesis chamber, and a mixer 107 is provided in the mixing chamber; the temperature control device 2 has a placement groove 2021 for placing the microfluidic chip 1 and a temperature control mechanism 201, and the temperature control mechanism 201 is used to control the temperature of the microfluidic chip 1 located in the placement groove 2021. When the microfluidic chip 1 performs DNA synthesis operation, precise temperature control can improve the reaction efficiency. At the same time, precise temperature control helps to ensure the consistency and integrity of the product, effectively reduce deviation, and improve the accuracy of synthesis.

[0047] Through the above technical solution, the synthesis chamber, mixing chamber and assembly chamber provided inside the microfluidic chip 1 can provide support for the synthesis of long-chain DNA without the need for transfer between different devices, thereby avoiding the alignment error caused by chip movement, reducing manual participation, reducing the mismatch rate and improving the overall efficiency and success rate of DNA synthesis. That is, a microelectrode 106 is provided in the synthesis chamber, and the reagent for synthesizing oligonucleotides is injected into the synthesis chamber through the liquid inlet 1011, and then directly contacts the microelectrode 106 in the synthesis chamber. The microelectrode 106 is controlled to start by the signal control circuit 4. After multiple rounds of synthesis reactions, the desired oligonucleotide array is obtained, and then the cleavage agent is injected into the synthesis chamber, and the microelectrode is selected according to the sequence design. 106 is energized to generate voltage, produce electrochemical reaction, and then break the chemical bond between the oligonucleotide and the electrode, cleave the oligonucleotide, and then inject the assembly reagent into the synthesis chamber through the liquid inlet hole 1011, and push the mixture containing the cleaved oligonucleotide in the synthesis chamber into the mixing chamber, and mix the cleaved oligonucleotide mixture and the assembly reagent by the mixer 107 arranged in the mixing chamber so that the two can be evenly mixed. Finally, the evenly mixed mixture is injected into the assembly chamber through the microchannel, and the microfluidic chip 1 is heated by the temperature control device 2 to provide a good environmental temperature for the assembly of long-chain DNA and improve the assembly efficiency. After the assembly is completed, the mixed solution containing the long-chain DNA is discharged from the liquid outlet 1012.

[0048] In some embodiments, reference Figures 5 to 8As shown, the microfluidic chip 1 includes a sealing plate 101, a reaction plate 102, a driving plate 103 and a bottom plate 104 which are fixedly connected in sequence, wherein the sealing plate 101 is provided with a liquid inlet hole 1011 and a liquid outlet hole 1012, the number of the liquid inlet holes 1011 is multiple, and the multiple liquid inlet holes 1011 are all connected to a liquid inlet tube, and one end of each liquid inlet tube away from the liquid inlet hole 1011 extends into a reagent bottle required for synthesis, and in the present application, a peristaltic pump can be used to extract the reagent in the reagent bottle and pass through the liquid inlet hole 1011 is injected into the synthesis chamber. Similarly, the liquid outlet 1012 is connected to a liquid outlet tube, and the liquid outlet tube is connected to a peristaltic pump, so as to extract the invalid liquid generated in the synthesis process (when the oligonucleotide array is synthesized in the synthesis chamber with the help of the microelectrode 106, a number of related reagents are needed. When the synthesis is completed, a large amount of flushing agent needs to be injected into the synthesis chamber through the liquid inlet 1011 and discharged from the liquid outlet 1012 to drain the residual liquid and reduce the influence of the synthesis reagents on the subsequent steps).

[0049] In some embodiments, reference Figures 5 to 8 As shown, the microfluidic chip 1 includes a sealing plate 101, a reaction plate 102, a driving plate 103 and a bottom plate 104 which are fixedly connected in sequence, wherein the synthesis chamber, the mixing chamber, the assembly chamber, the microchannel and the drainage channel are formed by combining the sealing plate 101, the reaction plate 102 and the driving plate 103, that is, the reaction plate 102 is provided with a synthesis through hole 1021, a mixing through hole 1022 and an assembly through hole 1023, and the synthesis through hole 1021 and the mixing through hole 1022 are connected to each other. 022 are connected by a flow channel 1024, the mixing through hole 1022 and the assembly through hole 1023 are connected by a flow channel 1024, and the end of the assembly through hole 1023 away from the flow channel is provided with a discharge channel 1025, and the driving plate 103 is provided with a synthesis tank 1031, a mixing tank 1032 and an assembly tank 1033, the synthesis tank 1031 is provided with a microelectrode 106, the mixing tank 1032 is provided with a mixer 107, and the synthesis tank 1031 and the mixing tank 1032 are connected by a through groove 1034, the mixing groove 1032 and the assembly groove 1033 are connected by a through groove 1034, and the end of the assembly groove 1033 away from the through groove 1034 is provided with a discharge groove 1035. After the sealing plate 101, the reaction plate 102 and the driving plate 103 are fixedly connected, the sealing plate 101, the synthesis through hole 1021 and the synthesis groove 1031 form a synthesis chamber, and the sealing plate 101, the mixing through hole 1022 and the mixing groove 103 2 forms a mixing chamber, the sealing plate 101, the assembly through hole 1023 and the assembly groove 1033 form an assembly chamber, the sealing plate 101, the flow channel and the through groove 1034 form a microchannel, the sealing plate 101, the discharge channel 1025 and the discharge groove 1035 form a liquid discharge channel, the bottom plate 104 is fixedly connected to the lower surface of the driving plate 103, and the bottom plate 104 is provided with a signal control circuit 4 for controlling the microvalve 105, the microelectrode 106 and the mixer 107.

[0050] In some embodiments, a microvalve 105 is provided in both the microchannel and the drainage channel. The microvalve 105 includes a piezoelectric ceramic that has been treated with corrosion protection. Under the action of an electrical signal, the piezoelectric ceramic is deformed to block the microchannel and the drainage channel. The opening and closing of the microvalve 105 should be controlled according to the actual synthesis process. For example, when an oligonucleotide array is synthesized in a synthesis chamber with the help of a microelectrode 106, the microchannel between the synthesis chamber and the mixing chamber should be closed; after the synthesis of the oligonucleotide array is completed, when a large amount of flushing agent is used to flush the synthesis chamber, the mixing chamber and the assembly chamber, the microvalve 105 of the microchannel between the synthesis chamber and the mixing chamber, the microvalve 105 of the microchannel between the mixing chamber and the assembly chamber, and the microvalve 105 provided in the drainage channel should all be opened, so that the flushing agent can carry the residual synthesis reagent through the drainage channel and be discharged from the liquid outlet 1012.

[0051] In some embodiments, reference Figure 7 As shown, a heating electrode 3 is provided in the assembly chamber, and the number of the heating electrodes 3 is multiple. The heating electrodes 3 are connected to the signal control circuit 4, and the opening and closing of the heating electrodes 3 are controlled by the signal control circuit 4. The setting of the heating electrode 3 enables the assembly chamber to heat up quickly, providing a good ambient temperature for the assembly of long-chain DNA, so as to ensure that enzymes such as ligase can effectively catalyze the reaction. At the same time, multiple heating electrodes 3 can cover a larger area, ensuring that the temperature distribution in the entire chamber is uniform, avoiding local overheating or uneven cooling, and, in the present application, a temperature control device 2 is also provided, through which the temperature of the microfluidic chip 1 can be controlled as a whole. On the basis of the temperature control device 2, with the help of the heating electrode 3, the assembly chamber can be heated more quickly and efficiently, thereby shortening the reaction time and improving the assembly efficiency.

[0052] In some embodiments, reference Figure 7 As shown, a plurality of microfluidic walls 5 are arranged at intervals in the synthesis chamber, and a microfluidic channel 501 for liquid circulation is formed between two adjacent microfluidic walls 5. In the process of synthesizing the oligonucleotide array, the synthesis reagent flows in an orderly manner under the guidance of the microfluidic channel 501. Microelectrodes 106 are arranged on the bottom surface of the microfluidic channel 501, and each microfluidic channel 501 has a plurality of microelectrodes 106 on the bottom surface. The synthesis reagent flows through these microelectrodes 106 in sequence, and the microelectrodes 106 are energized to generate voltage, and then an electrochemical reaction, such as a redox reaction, occurs to activate or convert the synthesis reagent and promote the synthesis of the oligonucleotide array; at the same time, the microfluidic channel 501 is arranged in the synthesis chamber to accurately control the flow and reaction conditions of liquids such as the synthesis reagent, such as flow rate, temperature, reaction time, etc., thereby improving the synthesis efficiency.

[0053] In some embodiments, reference Figure 7As shown, the mixer 107 includes a piezoelectric ceramic piece. Under the action of the electrical signal of the signal control circuit 4, the piezoelectric ceramic piece generates high-frequency vibration, causing liquid disturbance, thereby mixing the cleaved oligonucleotide mixture and the assembly reagent, and making the two mixed evenly.

[0054] In some embodiments not shown in the figure, in the present application, ultrasonic mixing can also be used to mix the cleaved oligonucleotide mixture and the assembly reagent, that is, the mixer 107 includes an ultrasonic module; at the same time, air disturbance mixing can also be used to mix the cleaved oligonucleotide mixture and the assembly reagent, that is, air inlet holes and air outlet holes are added to the sealing plate 101, and mixing is performed by air flow disturbance.

[0055] In some embodiments, reference Figure 1 , Figure 3 and Figure 4 As shown, the temperature control device 2 includes a substrate 202, a cover plate 203 and a fixing component 204, and the temperature control mechanism 201 includes a heat dissipation component and a heating plate 2011, wherein a placement groove 2021 is provided on the substrate 202, the cover plate 203 is arranged in the placement groove 2021 and is placed above the microfluidic chip 1, the fixing component 204 is arranged on the upper surface of the substrate 202 and has an abutment member for abutting the cover plate 203, the heat dissipation component is arranged on the lower surface of the substrate 202, and an embedding groove 2022 for accommodating the heating plate 2011 is provided on the bottom surface of the placement groove 2021.

[0056] In some embodiments, when the microfluidic chip 1 performs oligonucleotide synthesis, the microfluidic chip 1 is stably placed in the placement groove 2021 and covered with the cover plate 203, so that the microfluidic chip 1 is placed in a relatively closed chamber, and the microfluidic chip 1 is placed in a relatively heat-insulating environment, so as to avoid the synthesis of oligonucleotides affected by temperature fluctuations as much as possible. During the synthesis process, according to the progress of the synthesis, the heating plate 2011 and the heat dissipation component are used to perform precise temperature control, and the computer control system can be combined to realize real-time monitoring and adjustment of the temperature, so as to maintain the uniformity and stability of the temperature during the synthesis process, optimize the reaction efficiency and sequence specificity, and obtain good synthesis results.

[0057] Through the above technical scheme, when the microfluidic chip 1 is used to synthesize oligonucleotides, the microfluidic chip 1 is placed in the placement groove 2021 and the cover 203 is covered on the top of the microfluidic chip 1, and then the cover 203 is abutted by the abutment member of the fixing component 204, so as to fix the microfluidic chip 1. When the microfluidic chip 1 performs the synthesis operation, the microfluidic chip 1 can be heated by the heating plate 2011 and the microfluidic chip 1 can be cooled by the heat dissipation component. The temperature requirement of the microfluidic chip 1 during the synthesis process is met by heating and cooling, so as to achieve the purpose of improving the reaction efficiency and reducing the occurrence of non-specific reactions.

[0058] In some embodiments, the heat dissipation component includes a water-cooled plate, a water pump, a radiator, a water tank and a fan 701; wherein the water-cooled plate is attached to the bottom surface of the substrate 202, a passage for liquid circulation is provided in the water-cooled plate, and the two ends of the passage are respectively a liquid inlet and a liquid outlet, the liquid inlet and the liquid outlet are respectively connected with a liquid inlet pipe and a liquid outlet pipe, the inlet of the water pump is connected to the water tank through a connecting pipe, the outlet of the water pump is connected to the liquid inlet pipe, the liquid outlet pipe is connected to the inlet of the radiator, and the outlet of the radiator is connected to the water tank through a drain pipe. When the microfluidic chip 1 needs to be cooled, the water pump is started, and the water in the water tank flows out of the water tank, flows through the water pump, the water-cooled plate and the radiator in turn, and then flows back to the water tank through the drain pipe to complete the water circulation. With the continuous circulation of the water, the temperature of the microfluidic chip 1 is reduced to the required temperature. At the same time, in order to maximize the heat dissipation effect of the radiator, fan blades can be installed on the radiator to improve the heat dissipation effect.

[0059] In some embodiments, the fixing assembly 204 includes a base, a pressure rod and a rubber pressure head, wherein a through groove 2024 is provided on the base and a screw 2044 is threadedly connected to the base, the screw 2044 passes through the through groove 2024, one end of the pressure rod is provided with a through hole for the screw 2044 to pass through, and a rubber pressure head is installed on the other end of the pressure rod, the rubber pressure head abuts against the cover plate 203, and a torsion spring is installed at the connection between the screw 2044 and the pressure rod, so that the rubber pressure head is always abutted against the cover plate 203 through the torsion force of the torsion spring, thereby fixing the microfluidic chip 1.

[0060] In some embodiments, reference Figure 1 , Figure 3 and Figure 4As shown, the fixing assembly 204 includes a column 2041, a sleeve 2042, a connecting rod 2043 and a screw 2044, wherein one end of the column 2041 is fixedly connected to the upper surface of the substrate 202, the sleeve 2042 is sleeved on the column 2041, and the connecting rod 2043 is fixedly connected to the side wall of the sleeve 2042. In order to prevent the sleeve 2042 from being separated from the column 2041, the column 2041 may include a column and a limiting cap threadedly connected to the upper surface of the column. The upper surface of the column may be provided with a threaded hole, and the limiting cap is provided with a screw 2044. The limiting cap plays a limiting role to prevent the sleeve 2042 from being separated from the column. At the same time, because the sleeve 2042 and the The columns 2041 are sleeved together, and the sleeve 2042 and the connecting rod 2043 can rotate with the column 2041 as the center. When the cover 203 needs to be fixed, the connecting rod 2043 can be rotated so that the connecting rod 2043 is above the cover 203, and then the screw 2044 is rotated so that the screw 2044 gradually abuts against the cover 203, thereby completing the fixation of the cover 203 and achieving the purpose of fixing the microfluidic chip 1. At the same time, in order to facilitate personnel to turn the screw 2044, a handle 8 can be fixedly connected to the rod head of the screw 2044, and by turning the handle 8, personnel can turn the screw 2044 more easily and conveniently.

[0061] In some embodiments, the number of the fixing components 204 is multiple, and the multiple fixing components 204 are arranged at intervals along the edge of the substrate 202, that is, Figure 1 , Figure 3 and Figure 4 As shown, the fixing assembly can be arranged at an angle position of the base plate 202 , and a plurality of fixing assemblies 204 can fix the cover plate 203 from a plurality of angles, so that the cover plate 203 is more firmly placed in the placement groove 2021 .

[0062] In some embodiments, reference Figure 1 , Figure 3 and Figure 4As shown, the heat dissipation component includes a heat dissipation plate 2012, wherein the heat dissipation plate 2012 is installed on the lower surface of the substrate 202, and the number of the heat dissipation plates 2012 is multiple, and the multiple heat dissipation plates 2012 are arranged at intervals along the length direction of the substrate 202, that is, during the synthesis process, according to the synthesis requirements, the ambient temperature needs to be lowered. At this time, the heating plate 2011 can be turned off, and the heat emitted from the heating plate 2011 is effectively transferred to the heat dissipation plate 2012 through the multiple heat dissipation plates 2012 arranged at intervals, and then the heat is dispersed into the air through the heat dissipation plate 2012, so as to achieve the purpose of lowering the temperature of the microfluidic chip 1. At the same time, in order to improve the heat dissipation The heat dissipation effect of the heat plate 2012 can be achieved by the fan 701, that is, the fan 701 is installed on the mounting frame 7, and the edge of the lower surface of the substrate 202 is fixedly connected with a support column 703 for connecting to the mounting frame 7. The mounting frame 7 and the substrate 202 are connected by the support column 703. The mounting frame 7 is provided with a plug hole 702 for the support column 703 to be plugged in. At the same time, the fan 701 is located below the heat dissipation plate 2012, and the air outlet of the fan 701 is set toward the heat dissipation plate 2012. The fan 701 can effectively increase the circulation speed of the air around the heat dissipation plate 2012 and thereby improve the heat dissipation effect of the heat dissipation plate 2012.

[0063] In some embodiments, reference Figure 1 , Figure 3 and Figure 4 As shown, a support frame 6 is further provided below the mounting frame 7, that is, the support frame 6 is used to support the mounting frame 7 and to lift the mounting frame 7, so that a certain distance is maintained between the air inlet of the fan 701 and the laboratory desktop, and an opening for air circulation is provided on the support frame 6, and the air flows freely through the opening to the air inlet of the fan 701, ensuring that the fan 701 can blow out airflow efficiently, thereby allowing the heat sink 2012 to dissipate heat efficiently.

[0064] In some embodiments, reference Figure 1 , Figure 3 and Figure 4 As shown, in order to make the support frame 6 have good stability, a rubber gasket can be provided on the bottom surface of the support frame 6. With the help of the friction force of the rubber gasket, the support frame 6 can be stably placed on the laboratory desktop to prevent the support frame 6 from sliding on the laboratory desktop. At the same time, the rubber material has a certain elasticity and can absorb small vibrations between the desktop and the support frame 6, thereby improving the stability of the entire structure.

[0065] In some embodiments, reference Figure 1 , Figure 3 and Figure 4As shown, in order to facilitate personnel to take the heating plate 2011, hand-gripping grooves 2023 can be provided at both ends of the embedding groove 2022 that are relatively arranged along the length direction of the substrate 202, and the hand-gripping grooves 2023 and the embedding groove 2022 are interconnected, that is, when taking the heating plate 2011, the personnel can insert their fingers into the hand-gripping grooves 2023, and then make the fingertips fit against the heating plate 2011, so that the personnel can lift the heating plate 2011 located in the embedding groove 2022 from both sides of the heating plate 2011, thereby realizing efficient taking of the heating plate 2011 embedded in the embedding groove 2022.

[0066] In some embodiments, reference Figure 1 , Figure 3 and Figure 4 As shown, a power line is connected to the heating plate 2011, and the power line needs to extend to the outside of the substrate 202 and be connected to a power source. Therefore, a through groove 2024 is provided on the substrate 202 for the power line of the heating plate 2011 to pass through, and the through groove 2024 is interconnected with the embedded groove 2022. The power line of the heating plate 2011 extends to the outside of the substrate 202 after passing through the embedded groove 2022 and the through groove 2024. Similarly, a wire groove can also be opened on the substrate 202 for the relevant wire harness on the microfluidic chip 1 to extend to the outside of the substrate 202, wherein the relevant wire harness includes but is not limited to, data transmission line: if the microfluidic chip 1 integrates a sensor or other data acquisition device, a data line (such as a USB line, a serial line) is required to transmit data to an external computer or a data recorder; control line: for a microfluidic chip 1 integrated with a micropump, a valve or other controllable element, a control line is required to receive a signal from an external controller to regulate the flow of the fluid or other operations on the chip.

[0067] In some illustrated embodiments, the cover plate 203 needs to be punched, so that a plurality of through holes are opened on the cover plate 203. Each through hole is opened according to actual needs. In order to meet the synthesis requirements of the microfluidic chip 1, the plurality of through holes should correspond to the plurality of liquid inlet holes 1011 and the plurality of liquid outlet holes 1012, respectively.

[0068] Instructions for use of the temperature control device: Before placing the microfluidic chip 1 in the placement groove 2021, the heating plate 2011 can be placed in the embedding groove 2022, and the power cord of the heating plate 2011 can be combed so that the power cord extends to the outside of the substrate 202 through the through groove 2024. Then, the microfluidic chip 1 is placed in the placement groove 2021. Similarly, the relevant wire harness connected to the microfluidic chip 1 can be combed. A corresponding wire groove can be opened on the substrate 202, and the relevant wire harness connected to the microfluidic chip 1 can be extended to the outside of the substrate 202 through the wire groove. Then, the cover plate 203 is covered above the microfluidic chip 1 and the cover plate 203 is fixed by the fixing component 204. 3 is fixed, so that the microfluidic chip 1 can be stably placed in the placement groove 2021. After the cover plate 203 is fixed, the required liquid pipeline and gas pipeline are connected to the interface of the microfluidic chip 1 (the cover plate 203 is provided with through holes for the liquid pipeline and the gas pipeline to pass through), so as to meet the synthesis requirements of the microfluidic chip 1. When the synthesis operation is performed, the temperature is adjusted according to the synthesis requirements through the heat dissipation component and the heating plate 2011. The appropriate temperature can reduce the occurrence of non-specific reactions and ensure that the sequence of the synthesized oligonucleotide is accurate. At the same time, the appropriate temperature regulation can accelerate the rate of these chemical reactions, thereby improving the efficiency of the reaction.

[0069] On the basis of the above technical solution, the second aspect of the present invention further provides a DNA synthesis method based on the multi-chamber microfluidic chip 1, wherein DNA is synthesized by the multi-chamber microfluidic chip 1, and the DNA synthesis method comprises the following steps:

[0070] Step 101, injecting a synthesis reagent into the synthesis chamber through the liquid inlet 1011, so that the synthesis reagent reacts with the energized microelectrode 106 to obtain a target oligonucleotide array;

[0071] Step 102, injecting a cleaning agent into the synthesis chamber through the liquid inlet hole 1011, and discharging the cleaning agent after cleaning through the liquid outlet hole 1012;

[0072] Step 103, injecting a lysing agent into the synthesis chamber through the liquid inlet 1011, electrifying the microelectrode 106 to generate a voltage according to the sequence design, generating an electrochemical reaction to break the chemical bond between the oligonucleotide array and the microelectrode 106, and lysing the oligonucleotide array;

[0073] Step 104, injecting the assembly reagent into the synthesis chamber through the liquid inlet 1011, so that the assembly reagent and the mixed solution containing the oligonucleotide flow into the mixing chamber through the microchannel between the synthesis chamber and the mixing chamber;

[0074] Step 105, mixing the assembly reagent and the mixed solution containing the oligonucleotide by the mixer 107;

[0075] Step 106, discharging the mixed assembly reagent and the mixed solution containing the oligonucleotide into the assembly chamber through the microchannel located between the mixing chamber and the assembly chamber to assemble the long-chain DNA, and performing temperature control through the temperature control mechanism 201 and / or the heating electrode 3;

[0076] Step 107 , after the assembly is completed, the mixed solution containing the long-chain DNA is discharged from the assembly chamber through the liquid outlet 1012 .

[0077] The specific synthesis steps are described as follows:

[0078] The reagents required for synthesizing the oligonucleotide array (monomer ATCG, deprotecting agent, capping agent, activator, oxidant, flushing agent, etc.) are injected into the synthesis chamber through the liquid inlet 1011, and then directly contact the microelectrode 106 exposed in the synthesis chamber, and then the microelectrode 106 is controlled to start by the signal control circuit 4, and then a round of synthesis reaction occurs. For example: the deprotecting agent is injected into the synthesis chamber through the liquid inlet 1011, and then the microelectrode 106 is selected according to the sequence design to generate voltage, and the acid is generated within the range of the microelectrode 106 to remove the protecting group at the 5' end, exposing the hydroxyl group for subsequent coupling reaction, and the microelectrode 106 disconnects the voltage; the monomer (such as T) and the activator are injected into the synthesis chamber for activation coupling, and the hydroxyl group exposed in the previous step is connected to the monomer (such as T); the capping agent is injected into the synthesis chamber through the liquid inlet tube to seal the unreacted 5'-hydroxyl group; the oxidant is injected into the synthesis chamber through the liquid inlet tube to obtain a stable pentavalent phosphate bond;

[0079] Next, the deprotecting agent is injected into the synthesis chamber through the liquid inlet 1011, and then the microelectrode 106 is energized to generate voltage according to the sequence design, and the acid is generated within the microelectrode 106 to remove the protecting group at the 5' end, exposing the hydroxyl group for the subsequent coupling reaction, and the microelectrode 106 is disconnected from the voltage; the monomer (such as A) and the activator are injected into the synthesis chamber through the liquid inlet 1011 for activation coupling, and the hydroxyl group exposed in the above step is connected to the monomer (such as A); the capping agent is injected into the synthesis chamber through the liquid inlet 1011 to block the unreacted 5'-hydroxyl group; the oxidant is injected into the synthesis chamber through the liquid inlet 1011 to obtain a stable pentavalent phosphate bond. After that, the relevant synthesis operations are repeated by analogy to obtain the desired oligonucleotide array.

[0080] After the synthesis is completed, a large amount of flushing agent is injected into the synthesis chamber through the liquid inlet 1011 and discharged from the liquid outlet 1012 to drain the remaining liquid (a large amount of flushing agent and the reagents required for synthesizing the oligonucleotide array remaining after the synthesis) to reduce the impact of the synthesis reagents on subsequent steps.

[0081] Lysis step: The lysing agent is injected into the synthesis chamber through the liquid inlet 1011, and directly contacts the microelectrode 106 array and the oligonucleotide on the microelectrode 106 exposed in the synthesis chamber, and the microvalve 105 between the synthesis chamber and the mixing chamber is closed. According to the sequence design, the microelectrode 106 is energized to generate voltage, and an electrochemical reaction is generated to break the chemical bond between the oligonucleotide and the electrode, thereby lysing the oligonucleotide.

[0082] Mixing step: close the microvalve 105 between the mixing chamber and the assembly chamber, open the microvalve 105 between the synthesis chamber and the mixing chamber, inject the assembly reagent into the synthesis chamber through the liquid inlet 1011, push the mixture containing the cleavage oligonucleotide in the synthesis chamber to the mixing chamber, and continue to inject until the reaction system required for the assembly reaction is met, then apply a signal to the piezoelectric ceramic piece in the mixing chamber through the signal control circuit 4, and the piezoelectric ceramic piece generates vibration to mix the mixture containing the cleavage oligonucleotide and the assembly reagent (assembly system).

[0083] Assembly steps: close the microvalve 105 of the liquid discharge channel, open the microvalve 105 between the synthesis chamber and the mixing chamber, inject the assembly reagent into the synthesis chamber and the mixing chamber through the liquid inlet pipe, and then push the assembly system in the mixing chamber to the assembly chamber. Turn on the temperature control mechanism 201 and the heating electrode 3, adjust the temperature according to the assembly conditions, and quickly heat up through the temperature control mechanism 201 and the heating electrode 3 to provide a good temperature basis for the assembly process, thereby improving the assembly efficiency. After the assembly is completed, the mixed solution containing the long-chain DNA can be extracted through the liquid outlet 1012.

[0084] Those skilled in the art should understand that the discussion of any of the above embodiments is merely illustrative and is not intended to imply that the scope of the present disclosure (including the claims) is limited to these examples. Within the scope of the present invention, the above embodiments or technical features in different embodiments may be combined, the steps may be implemented in any order, and there are many other variations of the different aspects of the present invention as described above, which are not provided in detail for the sake of simplicity.

[0085] The embodiments of the present invention 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 the present invention should be included in the protection scope of the present invention.

Claims

1. A multi-chamber microfluidic chip, It is characterized in that include: A microfluidic chip (1), comprising a synthesis chamber, a mixing chamber, an assembly chamber, a liquid inlet hole (1011), a liquid outlet hole (1012), a microvalve (105), a microelectrode (106) and a mixer (107), wherein the synthesis chamber and the mixing chamber are connected via a microchannel, the mixing chamber and the assembly chamber are connected via the microchannel, an end of the assembly chamber away from the mixing chamber is connected to a liquid discharge channel, the microchannel and the liquid discharge channel are both provided with the microvalve (105), the liquid inlet hole (1011) is connected to the synthesis chamber, the liquid outlet hole (1012) is connected to the liquid discharge channel, the microelectrode (106) is provided in the synthesis chamber, and the mixer (107) is provided in the mixing chamber; The temperature control device (2) comprises a placement groove (2021) for placing the microfluidic chip (1) and a temperature control mechanism (201), wherein the temperature control mechanism (201) is used to control the temperature of the microfluidic chip (1) located in the placement groove (2021).

2. The multi-chamber microfluidic chip according to claim 1, It is characterized in that The microfluidic chip (1) comprises a sealing plate (101), a reaction plate (102), a driving plate (103) and a bottom plate (104) which are fixedly connected in sequence; The sealing plate (101) is provided with the liquid inlet hole (1011) and the liquid outlet hole (1012); the reaction plate (102) is provided with a synthesis through hole (1021), a mixing through hole (1022) and an assembly through hole (1023); the synthesis through hole (1021) and the mixing through hole (1022) are connected via a flow channel (1024); the mixing through hole (1022) and the assembly through hole (1023) are connected via the flow channel (1024); and the assembly through hole (1023) is opposite to the flow channel. The driving plate (103) is provided with a discharge channel (1025) at the end thereof, a synthesis tank (1031), a mixing tank (1032) and an assembly tank (1033) are provided on the driving plate (103), the synthesis tank (1031) is provided with the microelectrode (106), the mixing tank (1032) is provided with the mixer (107), the synthesis tank (1031) and the mixing tank (1032) are connected via a through-groove (1034), and the mixing tank (1032) and the assembly tank (1033) are connected via the through-groove (1034). The end of the assembly groove (1033) away from the through groove (1034) is provided with a discharge groove (1035). After the sealing plate (101), the reaction plate (102) and the driving plate (103) are fixedly connected, the sealing plate (101), the synthesis through hole (1021) and the synthesis groove (1031) form the synthesis chamber, the sealing plate (101), the mixing through hole (1022) and the mixing groove (1032) form the mixing chamber, and the sealing plate (101), the assembly through hole (1023) and the The assembly groove (1033) forms the assembly chamber, the sealing plate (101), the flow channel and the through groove (1034) form the microchannel, the sealing plate (101), the discharge channel (1025) and the discharge groove (1035) form the liquid discharge channel, the bottom plate (104) is fixedly connected to the lower surface of the driving plate (103) and a signal control circuit (4) for controlling the microvalve (105), the microelectrode (106) and the mixer (107) is provided inside the bottom plate (104).

3. The multi-chamber microfluidic chip according to claim 2, It is characterized in that A heating electrode (3) is provided in the assembly chamber, the number of the heating electrodes (3) is plural, and the heating electrode (3) is connected to the signal control circuit (4).

4. The multi-chamber microfluidic chip according to claim 1, It is characterized in that A plurality of microfluidic walls (5) are arranged at intervals in the synthesis chamber, and a microfluidic channel (501) for liquid circulation is formed between two adjacent microfluidic walls (5), and the microelectrode (106) is provided on the bottom surface of the microfluidic channel (501).

5. The multi-chamber microfluidic chip according to claim 1, It is characterized in that The mixer (107) comprises a piezoelectric ceramic sheet.

6. The multi-chamber microfluidic chip according to claim 1, It is characterized in that The temperature control device (2) comprises a base plate (202), a cover plate (203) and a fixing component (204); The substrate (202) is provided with the placement groove (2021), the cover plate (203) is arranged in the placement groove (2021) and is placed above the microfluidic chip (1), and the fixing component (204) is arranged on the upper surface of the substrate (202) and has an abutment member for abutting the cover plate (203); The temperature control mechanism (201) comprises a heat dissipation component and a heating plate (2011); the heat dissipation component is arranged on the lower surface of the base plate (202); and an embedding groove (2022) for accommodating the heating plate (2011) is provided on the bottom surface of the placement groove (2021).

7. The multi-chamber microfluidic chip according to claim 6, It is characterized in that The fixing assembly (204) comprises a column (2041), a sleeve (2042), a connecting rod (2043) and a screw rod (2044); One end of the column (2041) is fixedly connected to the upper surface of the base plate (202), the sleeve (2042) is sleeved on the column (2041) and the connecting rod (2043) is fixedly connected to the side wall of the sleeve (2042), the abutment member includes the screw rod (2044), and the end of the connecting rod (2043) away from the sleeve (2042) is provided with a threaded hole for threaded connection with the screw rod (2044).

8. The multi-chamber microfluidic chip according to claim 6, It is characterized in that The heat dissipation assembly comprises a heat dissipation plate (2012), the heat dissipation plate (2012) being mounted on the lower surface of the substrate (202), the number of the heat dissipation plates (2012) being multiple, and the multiple heat dissipation plates (2012) being arranged at intervals along the length direction of the substrate (202); It also comprises a support frame (6), a mounting frame (7) and a fan (701); a support column (703) for connecting to the mounting frame (7) is fixedly connected to the edge of the lower surface of the base plate (202); a plug hole (702) for plugging the support column (703) is provided on the mounting frame (7); the fan (701) is installed on the mounting frame (7) and the air outlet of the fan (701) is arranged toward the heat sink (2012); the support frame (6) is placed on the lower surface of the mounting frame (7); and a rubber gasket is provided on the bottom surface of the support frame (6).

9. The multi-chamber microfluidic chip according to claim 6, It is characterized in that The embedding groove (2022) is provided with hand-gripping grooves (2023) at two opposite ends thereof arranged along the length direction of the base plate (202); the hand-gripping grooves (2023) are in communication with the embedding groove (2022); a through groove (2024) for passing a power cord of the heating plate (2011) is provided on the base plate (202); the through groove (2024) is in communication with the embedding groove (2022); A handle (8) is fixedly connected to the rod head of the screw rod (2044); There are a plurality of fixing components (204), and the plurality of fixing components (204) are arranged at intervals along the edge of the substrate (202).

10. A DNA synthesis method based on a multi-chamber microfluidic chip, It is characterized in that DNA is synthesized by a multi-chamber microfluidic chip according to any one of claims 1 to 9, wherein the DNA synthesis method comprises: Injecting a synthesis reagent into the synthesis chamber through the liquid inlet (1011) so that the synthesis reagent undergoes a synthesis reaction with the energized microelectrode (106), thereby obtaining a target oligonucleotide array; Injecting a cleaning agent into the synthesis chamber through the liquid inlet (1011), and discharging the cleaning agent after cleaning through the liquid outlet (1012); A cleavage agent is injected into the synthesis chamber through the liquid inlet (1011), and the microelectrode (106) is energized to generate a voltage according to the sequence design, thereby generating an electrochemical reaction to break the chemical bond between the oligonucleotide array and the microelectrode (106), thereby cleaving the oligonucleotide array; Injecting the assembly reagent into the synthesis chamber through the liquid inlet (1011), so that the assembly reagent and the mixed solution containing the oligonucleotide flow into the mixing chamber through the microchannel between the synthesis chamber and the mixing chamber; Mixing the assembly reagent and the mixed solution containing the oligonucleotide by the mixer (107); The mixed assembly reagent and the mixed solution containing the oligonucleotide are discharged into the assembly chamber through the microchannel located between the mixing chamber and the assembly chamber to assemble the long-chain DNA, and the temperature is controlled by the temperature control mechanism (201) and / or the heating electrode (3); After the assembly is completed, the mixed solution containing the long-chain DNA is discharged from the assembly chamber through the liquid outlet (1012).

Images