A DNA synthesis device

Abstract

The present invention relates to a DNA synthesis device, comprising: a liquid reservoir group, the liquid reservoir group including a plurality of liquid storage bottles; a syringe pump group, the syringe pump group including a plurality of syringe pumps, the input ends of the syringe pumps being connected to the corresponding liquid storage bottles via pipelines; a multi-channel switching valve, the output ends of the plurality of syringe pumps being connected to the input port via pipelines; a microfluidic chip, the input end of the microfluidic chip being connected to the output port of the multi-channel switching valve via pipelines; a waste liquid bottle, the waste liquid bottle being connected to the output end of the microfluidic chip via pipelines; and a PLC controller, the PLC controller being electrically connected to the syringe pump group and used to control the operation of each syringe pump and the multi-channel switching valve. The DNA synthesis device optimizes the internal circuit and control system of the device, utilizes syringe pumps to accurately control the amount of reagents and consumables, saves resources, and avoids cross contamination between liquids.

Description

Technical Field

[0001] The invention belongs to the field of DNA synthesis, and in particular relates to a DNA fragment synthesis device used for synthesizing DNA fragments. Background Art

[0002] Generally speaking, human beings have entered the post-gene era. Simply understanding the composition of gene sequences can no longer meet the needs of scientific development. In contrast, scientists have also achieved remarkable results in biology, medicine and other fields through artificial DNA synthesis technology. One of the current methods of studying gene sequences is to use DNA synthesis devices to automatically synthesize DNA target fragments for subsequent research.

[0003] A DNA synthesizer is an instrument that performs biosynthesis in a reaction vessel, automatically connecting amino acids or nucleotides according to the base sequence of the target sequence. However, because the reaction requires the circulation of multiple liquids, cross-contamination between liquid paths is prone to occur when the reaction is performed on a synthesis column. The precise amount of enzyme used cannot be accurately achieved, resulting in waste of enzymes. Failure to promptly dispose of waste liquid after the reaction can damage the instrument or cause environmental pollution, significantly impacting the machine's service life. The reaction places high demands on the machine's operation and operating environment. Mutual interference between electronic circuits also makes it difficult for the instrument to perform reactions in complex industrial environments. Furthermore, the wiring within the equipment is often lengthy and complex, making routine maintenance and modifications difficult. Summary of the Invention

[0004] In response to the above problems, the purpose of the present invention is to provide a DNA synthesizer that optimizes the internal circuit and control system of the device, uses a syringe pump to accurately control the amount of reagent consumables, and saves resources. Each syringe pump of the liquid injection device has a corresponding independent multi-channel switching valve channel, thereby avoiding cross contamination between liquids.

[0005] To achieve the above object, the present invention adopts the following technical solutions:

[0006] The present invention provides a DNA synthesis device, comprising:

[0007] A liquid reservoir group, comprising a plurality of liquid storage bottles for storing raw materials for DNA synthesis, and the liquid reservoir group is stored at low temperature;

[0008] A syringe pump group, comprising a plurality of syringe pumps, each syringe pump corresponding to one of the liquid storage bottles, wherein an input end of the syringe pump is connected to the corresponding liquid storage bottle via a pipeline;

[0009] A multi-channel switching valve, wherein the multi-channel switching valve includes a plurality of input ports and an output port, wherein the output ends of the plurality of syringe pumps are connected to the input ports via pipelines, and each syringe pump corresponds to the input port;

[0010] A microfluidic chip, wherein an input end of the microfluidic chip is connected to an output port of the multi-channel switching valve via a pipeline, and the microfluidic chip is used for synthesizing DNA fragments;

[0011] A waste liquid bottle, which is connected to the output end of the microfluidic chip through a pipeline and is used to recover waste liquid generated during the DNA fragment synthesis process;

[0012] A PLC controller, wherein the PLC controller and the injection pump group use the UART protocol and adopt the RS485 communication line for connection and communication, and are used to control the action of each injection pump. The PLC controller and the multi-channel switching valve also use the RS485 communication line for connection and communication, and are used to control the one-to-one connection between different input ports and output ports.

[0013] Furthermore, the DNA synthesis device also includes a three-way valve, wherein the first end of the three-way valve is connected to the output port of the multi-channel switching valve via a pipeline to transmit the passing liquid, the second end is connected to the waste liquid bottle via a pipeline to discharge the initialized waste liquid, and the third end is connected to the input end of the microfluidic chip via a pipeline for reaction.

[0014] Furthermore, the DNA synthesis device further includes a heating device, which is connected to the PLC controller through I / O control, and the PLC controller controls the heating device to heat the microfluidic chip.

[0015] Furthermore, the heating device also includes a temperature measuring resistor and a temperature sensor. The resistance of the temperature measuring resistor increases linearly with the increase of temperature within the measurement range. The temperature measuring resistor transmits the measured value to the temperature sensor through a transmission line. The temperature sensor connects the temperature sensor power line to the third terminal for series power supply. After filtering and rectification, the data is sent to the analog input module of the PLC controller through the temperature sensor data line to detect the reaction temperature in real time.

[0016] Furthermore, the heating device includes an aluminum sheet and a heating ceramic, the aluminum sheet is connected to the heating ceramic, and the aluminum sheet and the heating ceramic are located below the microfluidic chip for supporting and heating the microporous core chip.

[0017] Furthermore, the DNA synthesis device also includes a refrigeration device, which includes a heat dissipation fan, a cooling plate, a cooling fan, a transistor temperature control module and a power supply. The transistor temperature control module is used to set the required cooling temperature and control the cooling to reach the required cooling temperature. The cooling plate has one side for cooling and the other side for heating. The heat dissipation fan is used to cool the heating surface, and the cooling fan is used to blow the cooling capacity of the cooling surface to the liquid storage tank group.

[0018] Furthermore, the DNA synthesis device also includes a base, and also includes a first terminal, a second terminal and a third terminal. The pump data communication lines of the multiple injection pump groups are connected to the PLC controller through the first terminal, and the power lines of the multiple injection pump groups are connected to the power supply through the second terminal. One end of the multi-channel switching valve, three-way valve and heating device is connected to the PLC controller, and the other end is connected to the power supply through the third terminal.

[0019] Furthermore, the DNA synthesis device also includes a first DC relay, a second DC relay and a third DC relay, one end of the first DC relay is connected to the PLC controller, and the other end is connected to the power supply, one end of the second DC relay is connected to the PLC controller, and the other end is connected to the light strip, and is connected to the power supply through the light strip, one end of the third DC relay is connected to the heating device, and the other end is connected to the power supply through the third DC relay.

[0020] Furthermore, the DNA synthesis device also includes a base, and the liquid storage tank group, injection pump group, multi-channel switching valve, microfluidic chip and waste liquid bottle are all installed on the base, and the base is also provided with a plurality of heat dissipation holes.

[0021] Due to the adoption of the above technical solution, the present invention has the following advantages: the DNA synthesis device provided by the present invention greatly reduces the number of logic units by wiring and using electromagnetic valves and relays, has a simple combined structure, and is convenient for equipment installation and maintenance. The device optimizes the internal circuit and control system of the device through a programmable logic controller, uses a syringe pump to accurately control the amount of reagent consumables, and saves resources. Each syringe pump of the liquid injection device has a corresponding independent multi-channel switching valve channel, which avoids cross contamination between liquids. The reaction in the microfluidic chip makes the reaction more complete and the efficiency is improved. After the reaction is completed, the device is cleaned using a cleaning device to ensure the normal use of the instrument. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Various other advantages and benefits will become apparent to those skilled in the art upon reading the detailed description of the preferred embodiment below. The accompanying drawings are for illustration purposes only and are not to be considered as limiting the present invention. Throughout the drawings, the same reference numerals are used to denote the same components. In the drawings:

[0023] Figure 1 It is the liquid circuit of the DNA synthesis device;

[0024] Figure 2 It is the circuit loop of the DNA synthesis device;

[0025] Figure 3 This is the assembly structure diagram of the DNA synthesis device.

[0026] Description of reference numerals:

[0027] 1-Liquid reservoir group, 2-Syringe pump group, 3-Multi-channel switching valve, 4-Three-way valve, 5-Waste liquid bottle, 6-Microfluidic chip, 7-Thermocouple resistor, 8-Ceramic heating plate, 9-PLC controller, 11-Temperature sensor, 12-Terminal block, 13-DC relay, 14-Refrigeration device, 15-Base, 121-First terminal block, 122-Second terminal block, 123-Third terminal block, 131-First DC relay, 132-Second DC relay, 133-Third DC relay, 141-Cooling fan, 142-Refrigeration plate, 143-Refrigeration fan, 144-Transistor temperature control module, 145-Power supply. DETAILED DESCRIPTION

[0028] Exemplary embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. Although exemplary embodiments of the present invention are shown in the accompanying drawings, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments described herein. Rather, these embodiments are provided to enable a more thorough understanding of the present invention and to fully convey the scope of the present invention to those skilled in the art.

[0029] An embodiment of the present invention provides a DNA synthesis device, comprising a liquid reservoir assembly, a syringe pump assembly, a multi-channel switching valve, a three-way valve, a microfluidic chip, a waste liquid bottle, and a PLC controller. The liquid reservoir assembly includes several liquid storage bottles for storing raw materials for DNA synthesis; the syringe pump assembly includes several syringe pumps, each corresponding to one liquid storage bottle, the input end of each syringe pump being connected to the liquid storage bottle via a pipeline; the multi-channel switching valve includes several input ports and one output port, the output ends of several syringe pumps being connected to the input ports via a pipeline, each syringe pump corresponding to one input port; the input end and the output port of the microfluidic chip are connected via a pipeline; the waste liquid bottle is connected to the output end of the microfluidic chip via a pipeline for collecting waste liquid generated during the DNA synthesis process; the PLC controller is connected to the syringe pump assembly via RS485 communication for controlling the operation of each syringe pump, and the PLC controller is connected to the multi-channel switching valve via RS485 communication for controlling the connection between different input ports and output ports. The DNA synthesis device optimizes the internal circuit and control system of the device through a programmable logic controller (PLC), and uses a syringe pump to accurately control the amount of reagents and consumables, saving resources. Each syringe pump of the liquid injection device has a corresponding independent multi-channel switching valve channel, avoiding cross-contamination between liquids. The reaction in the microfluidic chip makes the reaction more complete and the efficiency is improved. After the reaction is completed, the equipment is cleaned using a cleaning device to ensure the normal use of the instrument and extend the service life of the machine.

[0030] Example 1

[0031] An embodiment of the present invention provides a DNA synthesis device, comprising a base and, mounted on the base 15, a liquid reservoir assembly 1, a syringe pump assembly 2, a multi-channel switching valve 3, a three-way valve 4, a microfluidic chip 6, a waste liquid bottle 5, and a PLC controller 6. To facilitate heat dissipation, the base is also provided with a plurality of heat dissipation holes.

[0032] The liquid reservoir assembly 1 includes six liquid storage bottles, four of which are used to hold the bases and enzymes for DNA synthesis, one for a detergent, and one for a deprotection agent. The six liquid storage bottles are connected to the injection pump assembly 2 via different pipelines.

[0033] The syringe pump assembly includes six syringe pumps, each corresponding to a liquid storage bottle. The syringe pump input is connected to the corresponding liquid storage bottle via a pipeline. The syringe pump assembly 2 can precisely control the volume of liquid dispensed each time, with the flow rate and flow velocity displayed before each injection to ensure reaction accuracy and avoid unnecessary waste. The liquid in the liquid reservoir assembly 1 is transferred to the syringe pump via an infusion pipeline. The syringe pump then sequentially transfers the liquid to the multi-channel switching valve 3, where the corresponding pipeline inputs the corresponding liquid.

[0034] The multi-channel switching valve 3 includes six input ports and one output port, and the output ends of the six injection pumps are connected to the corresponding input ports through pipes; the input end and the output port of the microfluidic chip 6 are connected to each other through a pipeline, and each of the input port and output port is an inverted cone joint, and each of the inverted cone joints has an anti-backflow function, and liquid will flow out only when it is under pressure.

[0035] The input and output ports of the microfluidic chip 6 are connected via a pipeline. Different liquid storage bottles provide the liquid required for DNA synthesis to the microfluidic chip 6 via the syringe pumps and the switching valve. The PLC controller 9 is electrically connected to the syringe pump assembly 2 to control the operation of each syringe pump. The PLC controller 9 is also electrically connected to the multi-channel switching valve 3 to control the communication between the different input ports and the output ports.

[0036] During the DNA synthesis process, the groups in the microfluidic chip 6 are first cleaned by the cleaning liquid, and then the PLC controller 9 controls different liquid storage bottles to provide the required bases and biological enzymes. Finally, the PLC controller 9 controls the supply of deprotection agent to the microfluidic chip 6 to fix the synthesized DNA.

[0037] The DNA synthesis device optimizes the internal circuit and control system of the device through the programmable logic controller 9, and uses the injection pump to accurately control the amount of reagent consumables to save resources. Each injection pump of the liquid injection device has a separate multi-channel switching valve 3 corresponding to each channel, which avoids cross-contamination between liquids. The reaction in the microfluidic chip 6 makes the reaction more complete and the efficiency is improved. After the reaction is completed, the equipment is cleaned using a cleaning device to ensure the normal use of the instrument.

[0038] The DNA synthesizer includes three terminals 12, namely a first terminal 121, a second terminal 122, and a third terminal 123. The first terminal 121 is used for data signal communication between the multiple syringe pump groups 2 and the PLC controller 9. The second terminal 122 is used for connecting the multiple syringe pump groups 2 to a power supply 10. The PLC controller 9 and the syringe pump groups 2 are connected to the power supply 10 via the third terminal 123. The power supply 10 is used to power the syringe pump groups 2 and the PLC controller 9.

[0039] Each syringe pump in syringe pump set 2 uses two data transmission lines, for a total of twelve data lines. The data transmission lines are divided into A-end and B-end. The six A-ends of the data transmission lines are connected in series through the first wiring terminal 121, and the six B-ends are connected in series. The two main data lines led out from the other side of the first wiring terminal are connected to the PLC controller 9, completing the data transmission wiring of syringe pump set 2. The PLC controller 9 is an S1200 PLC controller.

[0040] Each injection pump of the injection pump group 2 uses two power cords, for a total of twelve power cords. The power cords are divided into positive and negative poles. 24V DC (direct current) is required. The six positive poles of the power cords are connected in series through the second terminal 122, and then the six negative poles are connected in series. The two pump power buses drawn out from the other side of the second terminal 122 are connected to the 51DC24V industrial DC power supply 10 to complete the wiring of the power supply part of the injection pump group 2.

[0041] The switching valve data line that controls the operation of the multi-channel switching valve 3 is directly connected to the PLC controller 9 through a conversion connector to complete the data part wiring of the multi-functional switching valve 3. The power line of the multi-functional switching valve 3 is directly powered in series through the third terminal 123.

[0042] In order to initialize the pipeline and discharge the air in the pipeline, the DNA synthesis device also includes a three-way valve 4, the first end of the three-way valve 4 is connected to the output port of the multi-channel switching valve 3 by a pipeline, the second end is connected to the waste liquid bottle 5 by a pipeline, and the third end is connected to the microfluidic chip 6 by a pipeline. Before providing the synthetic liquid to the microfluidic chip 6, the air in the pipeline is first emptied by connecting the three-way valve 4 to the waste liquid bottle 5, so as to accurately control the amount of reagents and consumables using an injection pump, save resources, and ensure that the reaction proceeds smoothly. One end of the three-way valve 4 is connected to the I / O control of the PLC controller 9, and the conduction and closing of different channels of the three-way valve 4 are controlled by the PLC controller 9. The other end of the three-way valve 4 is connected to the power supply 10 through the third terminal 123.

[0043] The DNA synthesis device also includes a heating device, which comprises an aluminum sheet and a ceramic heating sheet 8. The microfluidic chip 6 is placed on the aluminum sheet and ceramic heating sheet 8. One end of the ceramic heating sheet 8 is connected to the I / O control of the PLC controller 9, and the other end is electrically connected to the power supply 10 via the third terminal 123. The PLC controller 123 controls the heating temperature of the ceramic heating sheet 8. The heating device also includes a temperature measuring resistor 7 and a temperature sensor 11. The resistance of the temperature measuring resistor 7 increases linearly with increasing temperature within the measurement range. The temperature measuring resistor 7 transmits the measured value to the temperature sensor 11 via a transmission line. The temperature sensor 11 connects the temperature sensor power line to the third terminal 123 for series power supply. After filtering and rectification, the data is sent to the analog input module of the PLC controller 9 via the temperature sensor data line, completing the temperature control system.

[0044] When the device is running, a running light strip 16 will flash to indicate that the system is running. One end of the light strip is electrically connected to the PLC controller 9, and the other end is connected to the power supply through the third terminal 123. The PLC controller 9 controls the operation of the light strip 16.

[0045] The DNA synthesis device also includes three DC relays 13, namely a first DC relay 131, a second DC relay 132 and a third DC relay 133. One end of the first DC relay 131 is connected to the PLC controller 9, and the other end is connected to the power supply 10. One end of the second DC relay 132 is connected to the PLC controller 9, and the other end is connected to the light strip 16. The light strip 16 is connected to the power supply 10. One end of the third DC relay 133 is connected to the heating device, and the other end is connected to the power supply 10 through the third DC relay 123.

[0046] The DNA synthesis device also includes a refrigeration unit 14, which is used to cool the liquid storage tank group 1, allowing the refrigeration system to maintain a low temperature for a long time. The refrigeration unit 14 includes a cooling fan 141, a cooling plate 142, a cooling fan 143, a transistor temperature control module 144, and a power supply 145. The transistor temperature control module 144 is used to control the cooling temperature of the cooling plate 142. The cooling plate 142 cools on one side and heats on the other side. The cooling fan 141 is used to cool the heating surface, and the cooling fan 143 is used to blow the cooling water toward the liquid storage tank group 1. In addition, the transistor temperature control module 144 can manually adjust the temperature of the refrigeration system to maintain a constant temperature, which is conducive to the reaction.

[0047] When assembling the DNA, the injection pump group 2 and the refrigeration module are arranged up and down, and the three-way valve 4 and the multi-channel switching valve 3 are arranged up and down. This distribution is conducive to shortening the length of the pipeline, so that the dead volume in the pipeline will be reduced, which is more conducive to the full progress of the reaction and saves materials, avoiding waste. The microfluidic chip 6 is above the waste liquid pool, and a large fan is installed in the equipment casing to cool all modules in the machine. Small holes are also punched on the casing, which is also conducive to cooling the equipment. In addition, the PLC controller 9 and relays, etc. are all on the back side of the injection pump to facilitate wiring, avoid unnecessary equipment arrangement, and cause confusion in the lines of the equipment, etc.

[0048] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. A DNA synthesis device, characterized in that: include: A liquid reservoir group, comprising a plurality of liquid storage bottles for storing raw materials for DNA synthesis, and the liquid reservoir group is stored at low temperature; A syringe pump group, comprising a plurality of syringe pumps, each syringe pump corresponding to one of the liquid storage bottles, wherein an input end of the syringe pump is connected to the corresponding liquid storage bottle via a pipeline; A multi-channel switching valve, wherein the multi-channel switching valve includes a plurality of input ports and an output port, wherein the output ends of the plurality of syringe pumps are connected to the input ports via pipelines, and each syringe pump corresponds to the input port; A microfluidic chip, wherein an input end of the microfluidic chip is connected to an output port of the multi-channel switching valve via a pipeline, and the microfluidic chip is used for synthesizing DNA fragments; A waste liquid bottle, which is connected to the output end of the microfluidic chip through a pipeline and is used to recover waste liquid generated during the DNA fragment synthesis process; A PLC controller, wherein the PLC controller is connected and communicated with the injection pump group using a communication line, and is used to control the action of each injection pump. The PLC controller is also connected and communicated with the multi-channel switching valve using a communication line, and is used to control the one-to-one communication between different input ports and output ports; The system further comprises a three-way valve, wherein a first end of the three-way valve is connected to an output port of the multi-channel switching valve via a pipeline to transmit past liquid, a second end is connected to the waste liquid bottle via a pipeline to discharge initialized waste liquid, and a third end is connected to an input end of the microfluidic chip via a pipeline for reaction. The three-way valve and the multi-channel switching valve are arranged in an up-down manner to shorten the length of the pipeline. The microfluidic chip is above the waste liquid tank, and the PLC controller is on the rear side of the injection pump to facilitate wiring. The system also comprises a heating device, which is connected to the PLC controller for I / O control, and the PLC controller controls the heating device to heat the microfluidic chip. The heating device also includes a temperature measuring resistor and a temperature sensor. The resistance of the temperature measuring resistor increases linearly with the increase of temperature within the measurement range. The temperature measuring resistor transmits the measured value to the temperature sensor through a transmission line. The temperature sensor connects the temperature sensor power line to the third terminal for series power supply. After filtering and rectification, the data is sent to the analog input module of the PLC controller through the temperature sensor data line for real-time detection of the reaction temperature. The heating device includes an aluminum sheet and a heating ceramic, wherein the aluminum sheet is connected to the heating ceramic, and the aluminum sheet and the heating ceramic are located below the microfluidic chip and are used to support and heat the microfluidic chip; It also includes a refrigeration device, which includes a heat dissipation fan, a cooling plate, a cooling fan, a transistor temperature control module and a power supply. The transistor temperature control module is used to set the required cooling temperature and control the cooling to reach the required cooling temperature. The cooling plate has one side for cooling and the other side for heating. The heat dissipation fan is used to cool the heating surface, and the cooling fan is used to blow the cooling capacity of the cooling surface to the liquid storage tank group.

2. The DNA synthesis device according to claim 1, characterized in that It also includes a first wiring terminal, a second wiring terminal and a third wiring terminal. The pump data communication lines of the multiple injection pump groups are connected to the PLC controller through the first wiring terminal, and the power lines of the multiple injection pump groups are connected to the power supply through the second wiring terminal. One end of the multi-channel switching valve, three-way valve and heating device is connected to the PLC controller, and the other end is connected to the power supply through the third wiring terminal.

3. The DNA synthesis device according to claim 1, characterized in that It also includes a first DC relay, a second DC relay and a third DC relay, one end of the first DC relay is connected to the PLC controller, and the other end is connected to the power supply, one end of the second DC relay is connected to the PLC controller, and the other end is connected to the light strip, and is connected to the power supply through the light strip, one end of the third DC relay is connected to the heating device, and the other end is connected to the power supply through the third DC relay.

4. The DNA synthesis device according to claim 1, characterized in that: It also includes a base, on which the liquid storage tank group, injection pump group, multi-channel switching valve, microfluidic chip and waste liquid bottle are all installed, and the base is also provided with a plurality of heat dissipation holes.

Images