Full-automatic high-flux water bath PCR (Polymerase Chain Reaction) thermal cycler and control method thereof
The design of a fully automated high-throughput water bath PCR thermal cycler solves the problems of low amplification efficiency and poor temperature control accuracy of existing water bath PCR instruments, achieving efficient and accurate sample processing and consistency of experimental results.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-31
- Publication Date
- 2026-04-03
AI Technical Summary
Existing water bath PCR instruments have low amplification efficiency, poor temperature control accuracy, and problems such as uneven water temperature distribution, slow cooling speed, and risk of dry burning, making it difficult to meet the needs of high-end amplification.
Design a fully automated high-throughput water bath PCR thermal cycler, which employs a parallel water bath, heat dissipation components, and horizontal and vertical drive units, combined with heating and heat dissipation control, to achieve automated sample processing and precise temperature control.
This improved the throughput and temperature control accuracy of the water bath PCR instrument, shortened the experimental time, and significantly improved experimental efficiency and the consistency of results.
Smart Images
Figure CN121780315A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of PCR amplification technology, and in particular to a fully automated high-throughput water bath PCR thermal cycler and its control method. Background Technology
[0002] Currently available water bath PCR instruments are primarily low-throughput devices with limited functionality, making them unsuitable for rapid PCR amplification of large numbers of samples. Traditional water bath PCR instruments typically feature three independent water baths, each with its own temperature controlled by a stainless steel heating element or a PTC ceramic heater. During the experiment, the operator must manually or semi-automatically transfer the reaction plates or tubes between the three baths according to a pre-set temperature cycling protocol to complete the PCR amplification. This method is cumbersome and significantly reduces amplification efficiency.
[0003] Meanwhile, the existing temperature control method for water bath PCR equipment is to directly heat the water from the bottom of the bath to raise the water temperature to the set temperature. This method has the following obvious shortcomings: First, this heating method leads to localized high temperatures at the bottom, and the water in the bath lacks a mixing and stirring device, resulting in uneven water temperature distribution, affecting the consistency of sample amplification results, and the amplification efficiency may vary in different locations; Second, the water bath with the set annealing temperature usually relies solely on natural cooling or basic refrigeration to lower the temperature, lacking a precise cooling control system, resulting in slow cooling speed and low temperature control accuracy, making it difficult to meet the requirements of high-end amplification programs such as Touchdown PCR that require precise temperature gradient changes; Third, there is a lack of a linkage control mechanism between water level and temperature. When the water level is too low, it may cause the risk of dry burning, or even lead to equipment temperature control failure, which not only threatens equipment safety but also affects experimental stability and the accuracy of results.
[0004] Therefore, a fully automated high-throughput water bath PCR thermal cycler and its control method were developed to solve the above problems. Summary of the Invention
[0005] This invention proposes a fully automated high-throughput water bath PCR thermal cycler and its control method to solve the problems of low amplification efficiency and poor temperature control accuracy of existing water bath PCR instruments.
[0006] The present invention achieves the above objectives through the following technical solutions: On one hand, the present invention provides a high-throughput water bath PCR thermal cycler, comprising: The thermal circulation unit includes at least three water baths and a heat dissipation component. The water baths are arranged in parallel, and the top of each water bath is open. Each water bath includes a water tank, a heating component, a temperature detection module, and a water inlet. The heating component is located inside the water tank. The temperature detection module is used to detect the water temperature inside the water tank of each water bath. The heat dissipation component is used to dissipate heat and cool down at least one water bath. The water inlet is used to connect to an external water source. Loading unit; the loading unit is used for batch loading of reaction plates; A horizontal drive unit is used to drive the loading unit to move horizontally to directly above each water bath tank; The vertical drive unit is used to drive the loading unit to descend into the interior of each water bath tank, or to drive the loading unit to rise and leave the interior of each water bath tank. The control unit is connected to the heating components, horizontal drive unit, vertical drive unit temperature detection module and heat dissipation components in each water bath.
[0007] Furthermore, each water bath is equipped with a return water inlet at the top. The heating components include heating pipes and heating rods. The heating pipes are horizontally installed at the bottom of the water tank, and the heating rods are installed inside the heating pipes. Several guide spray holes are installed on the top of the heating pipes, and a water inlet is installed at the bottom of the heating pipes. The water inlet and the return water inlet are connected by a circulation pump.
[0008] Furthermore, the heat dissipation component includes a fan, heat pipes, and heat dissipation fins. The inlet end of the heat pipe is connected to the outlet of each circulation pump, and the outlet end of the heat pipe is connected to the inlet of the corresponding water bath. An inlet water temperature detection module is provided at the inlet. The fan is set close to the inner side of the heat pipe, and the heat dissipation fins are set close to the outer side of the heat pipe. The fan, the inlet water temperature detection module, and the control unit are connected.
[0009] Furthermore, the horizontal drive unit includes a horizontal guide rail, a first stepper motor, a first slider, a first synchronous belt, and a first synchronous pulley. The front and rear ends of the horizontal guide rail are respectively provided with first synchronous pulleys. The two first synchronous pulleys are connected by the first synchronous belt. The first stepper motor is driven by one of the first synchronous pulleys. The first slider is supported by the loading unit. The first slider is fixedly connected to the first synchronous belt. The first slider is slidably connected to the horizontal guide rail.
[0010] Furthermore, it also includes a cover assembly, which includes a high-temperature cloth and a cover frame. The high-temperature cloth covers the outside of the first synchronous belt and moves with the first synchronous belt. The high-temperature cloth is provided with a cover frame that is the same size as the opening of each water bath. One end of the cover frame is fixedly connected to the first slider.
[0011] Furthermore, the vertical drive unit includes a vertical guide rail, a second stepper motor, a second slider, a second synchronous belt, a second synchronous pulley, a counterweight, a guide rod, and a pulley assembly. The upper and lower ends of the vertical guide rail are respectively equipped with second synchronous pulleys, which are connected by a second synchronous belt. The second stepper motor is driven by one of the second synchronous pulleys. The second slider is connected to the loading unit and fixedly connected to the second synchronous belt. The second slider is slidably connected to the vertical guide rail. A guide rod connects the upper and lower ends of the vertical guide rail. The counterweight passes through the guide rod and is fixedly connected to the second synchronous belt. When the second slider moves to the upper end of the vertical guide rail, the counterweight is located at the lower end of the vertical guide rail. The pulley assembly is located at the top of the vertical guide rail, and the second slider and the counterweight are connected by a suspension rope mounted on the pulley assembly.
[0012] Furthermore, the loading unit includes a water bath basket and a water bath basket bracket. The water bath basket bracket is fixedly connected to the second slider. The water bath basket bracket includes a spring, a water tank cover, and a bracket connected sequentially from top to bottom. The water tank cover fits into the top opening of each water bath. The upper end of the water tank cover is connected to the second slider through at least one spring. Horizontal guide grooves are provided on both sides of the bottom of the bracket. Horizontal slide rails that are adapted to and engaged with the guide grooves are provided on both sides of the top of the water bath basket.
[0013] Furthermore, it also includes a level gauge, which is installed on the outer wall of each water tank and is used to measure the water level inside the tank.
[0014] Furthermore, it also includes a frame assembly, the upper and lower parts of which are separated by a water bath frame. The water bath frame has openings corresponding to the top openings of each water bath. The heat circulation unit is located in the lower part of the frame assembly, while the horizontal drive unit and the vertical drive unit are both located in the upper part of the frame assembly.
[0015] On the other hand, the present invention also provides a control method for the aforementioned high-throughput water bath PCR thermal cycler, comprising: Load the reaction plate into the loading unit; Obtain the PCR amplification experimental procedure set by the user; The user-defined PCR amplification experimental workflow is initiated. The PCR amplification experimental workflow execution process is as follows: The horizontal drive unit is controlled to move the loading unit directly above the target water bath tank; Determine whether the liquid level in each water bath has reached the upper liquid level. If not, replenish the liquid to the upper liquid level. If so, issue the target temperature for each water bath and control the heating components in each water bath to heat it. Once the temperature of each water tank reaches the starting temperature of the experiment, the remaining water tanks are subjected to timed temperature control, while the target water tank continues to be heated. When the real-time temperature of the target water bath reaches the target temperature, the vertical drive unit is controlled to lower the loading unit into the target water bath and fully immerse it in the water. Then, the loading unit is controlled to make small up-and-down swaying motions in the water. The heating and heat dissipation components are controlled to keep the real-time temperature of the target water bath at the target temperature. At the same time, the temperature of the next target water bath is controlled to be heated to the target temperature. Once the reaction in the target water bath is complete and the next target water bath reaches the target temperature, the vertical drive unit is controlled to lift the loading unit, and the horizontal drive unit is controlled to move the loading unit to the next target water bath, and the above experimental procedure is repeated.
[0016] The beneficial effects of this invention are as follows: The present invention proposes a fully automated high-throughput water bath PCR thermal cycler and its control method, which improves the processing throughput of the water bath PCR instrument, shortens the experimental time of the water bath PCR instrument, realizes the fully automated experimental process, significantly improves the experimental efficiency of the water bath PCR instrument, and greatly improves the temperature control accuracy during the experimental process. Attached Figure Description
[0017] Figure 1 This is a schematic diagram of the internal assembly structure of a fully automated high-throughput water bath PCR thermal cycler according to an embodiment of this application; Figure 2 This is a schematic diagram of the thermal cycling unit in an embodiment of this application; Figure 3 This is a schematic diagram of the loading unit in an embodiment of this application; Figure 4 This is a schematic diagram of the structure of the water bath basket in the embodiments of this application; Figure 5 This is a schematic diagram of the structure of the horizontal drive unit in the embodiments of this application; Figure 6 This is a schematic diagram of the vertical drive unit in an embodiment of this application; Figure 7 This is a schematic diagram of the structure of the fabric cover assembly in an embodiment of this application; Figure 8 This is a schematic diagram of the external structure of a fully automated high-throughput water bath PCR thermal cycler according to an embodiment of this application; Figure 9 This is a schematic diagram of the structure of the water bath tank in the embodiments of this application; Figure 10 This is a schematic diagram of the heating component in an embodiment of this application; Figure 11 This is a schematic diagram of the heat dissipation component in an embodiment of this application; Figure 12 This is a schematic diagram of the structure of the framework component in the embodiments of this application; Figure 13 This is a schematic diagram of the temperature control process in an embodiment of this application; Figure 14 for Figure 3 A magnified schematic diagram of the structure of A in the middle.
[0018] In the diagram: 1-Leakage protection switch; 2-Power switch; 3-Equipment door; 4-Water bath basket; 5-Overflow port; 6-High temperature water tank; 7-Low temperature water tank; 8-Ordinary water tank; 9-Reaction plate; 10-Reaction plate slot; 11-Water bath basket baffle; 12-Water bath basket hanger; 13-Horizontal drive unit; 14-First stepper motor; 15-First synchronous belt drive system; 151-First synchronous belt; 152-First synchronous pulley; 16-Horizontal guide rail; 17-First slider; 18-Cloth cover assembly; 19-High temperature cloth; 20-Cloth cover frame; 21-Vertical drive unit; 22-Counterweight; 23-Pulley block; 24-Second stepper motor; 25-Second synchronous belt drive system; 251-Second synchronous belt ; 252-Second Synchronous Wheel; 26-Vertical Guide Rail; 27-Guide Rod; 28-Heat Circulation Unit; 29-Return Water Inlet; 30-Water Inlet; 31-Level Gauge; 32-Idle Water Tank Assembly; 33-Heat Dissipation Component; 34-Circulation Pump; 35-Frame Assembly; 36-Profile Frame; 37-Screen Assembly; 38-Electrical Box Assembly; 39-Sliding Door Assembly; 40-Water Tank Cover; 41-Spring; 42-Positioning Bead; 43-Positioning Groove; 44-Side Plate; 45-Fixing Rod; 46-Mounting Plate; 47-Heating Pipe; 48-Heating Rod; 49-Guide Spray Hole; 50-Fan; 51-Heat Dissipation Box; 52-Water Inlet End of Heat Dissipation Pipe; 53-Water Outlet End of Heat Dissipation Pipe; 54-Water Inlet. Detailed Implementation
[0019] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. The components of the embodiments of the present invention described and shown in the accompanying drawings can generally be arranged and designed in various different configurations.
[0020] Therefore, the following detailed description of the embodiments of the invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the invention without inventive effort are within the scope of protection of the invention.
[0021] It should be noted that similar labels and letters in the following figures indicate similar items. Therefore, once an item is defined in one figure, it does not need to be further defined and explained in subsequent figures.
[0022] In the description of this invention, it should be understood that the terms "upper," "lower," "inner," "outer," "left," "right," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, or the orientation or positional relationship commonly used when the product of this invention is in use, or the orientation or positional relationship commonly understood by those skilled in the art. They are only used to facilitate the description of this invention and to simplify the description, and are not intended to indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this invention.
[0023] Furthermore, the terms "first," "second," etc., are used only to distinguish descriptions and should not be interpreted as indicating or implying relative importance.
[0024] In the description of this invention, it should also be noted that, unless otherwise explicitly specified and limited, terms such as "set" and "connection" should be interpreted broadly. For example, "connection" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium; it can be a connection within two components. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.
[0025] The specific embodiments of the present invention will now be described in detail with reference to the accompanying drawings.
[0026] like Figure 1 As shown, a fully automated high-throughput water bath PCR thermal cycler includes: The thermal circulation unit 28 includes at least three water baths and a heat dissipation component 33. The upper end of each water bath is open. Each water bath includes a water tank, a heating component, a temperature detection module, and a water inlet 54. The heating component is located inside the water tank. The temperature detection module is used to detect the water temperature inside the water tank of each water bath. The heat dissipation component 33 is used to dissipate heat and cool down at least one water bath. The water inlet 54 is used to connect to an external water source. Loading unit; the loading unit is used for batch loading of reaction plates; The horizontal drive unit 13 is used to drive the loading unit to move horizontally to directly above each water bath tank; The vertical drive unit 21 is used to drive the loading unit to descend vertically into the interior of each water bath, or to drive the loading unit to rise and leave the interior of each water bath. The control unit is connected to the heating components, horizontal drive unit 13, vertical drive unit 21 temperature detection module and heat dissipation component 33 in each water bath.
[0027] The thermal circulation unit 28 includes a high-temperature water tank 6, a low-temperature water tank 7, and a normal water tank 8 arranged sequentially. A heat dissipation component 33 is used to cool the normal water tank 8.
[0028] like Figure 3 As shown, the loading unit includes a water bath basket 4 and a water bath basket bracket 12. The water bath basket bracket 12 is fixedly connected to the second slider. The water bath basket bracket 12 includes a spring 41, a water tank cover 40, and a bracket connected sequentially from top to bottom. The water tank cover 40 fits into the top opening of each water bath. The upper end of the water tank cover 40 is connected to the second slider through at least one spring 41. Horizontal guide grooves are provided on both sides of the bottom of the bracket. Horizontal slide rails that are adapted to and engaged with the guide grooves are provided on both sides of the top of the water bath basket 4. The thermal circulation unit 28 also includes an idle water tank assembly 32, which is used to add or remove water baths according to experimental needs.
[0029] like Figure 4 As shown, the water bath basket 4 includes: a frame, the frame including two side plates 44, the two side plates 44 being located at opposite ends of the frame and arranged parallel to each other; and mounting plates 46, the two side plates 44 being connected by at least two mounting plates 46, the mounting plates 46 being spaced vertically apart, each of the two opposite surfaces of two adjacent mounting plates 46 having a row of reaction plate slots 10, the reaction plate slots 10 on adjacent mounting plates being vertically aligned, and the space between a pair of vertically aligned reaction plate slots 10 being used for the insertion of the upper and lower ends of a single reaction plate 9, the space between two adjacent mounting plates 46 forming a reaction plate mounting position. Water bath basket baffles 11 are provided on both sides of the frame, the water bath basket baffles 11 on both sides of the reaction plate mounting position.
[0030] like Figure 2 , Figure 9 and Figure 10 As shown in the figure, in one embodiment, each water bath is also provided with a return water inlet 29 at the top. The heating component includes a heating pipe 47 and a heating rod 48. The heating pipe 47 is horizontally arranged at the bottom of the water tank. The heating rod 48 is arranged inside the heating pipe 47. Several guide spray holes 49 are provided on the top of the heating pipe 47. The bottom of the heating pipe 47 is provided with a water inlet 30. The water inlet 30 and the return water inlet 29 are connected to the circulation pump 34 through a circulation pipeline.
[0031] like Figure 11In one embodiment, the heat dissipation component 33 is connected to the ordinary water tank 8. The heat dissipation component 33 includes a fan 50, a heat pipe, and heat dissipation fins. The heat pipe and heat dissipation fins are both installed inside the heat dissipation box 51. The water inlet 52 of the heat pipe is connected to the water outlet of the circulation pump 34, and the water outlet 53 of the heat pipe is connected to the water inlet 30 of the water bath. A return water temperature detection module is provided at the return water inlet 29. The fan 50 is arranged close to the inner side of the heat pipe, and the heat dissipation fins are arranged close to the outer side of the heat pipe. The fan 50, the return water temperature detection module, and the control unit are connected.
[0032] like Figure 5 As shown, in one embodiment, the horizontal drive unit 13 includes a horizontal guide rail 16, a first stepper motor 14, a first slider 17, a first synchronous belt 151, and a first synchronous pulley 152. The front and rear ends of the horizontal guide rail 16 are respectively provided with first synchronous pulleys 152. The two first synchronous pulleys 152 are connected by the first synchronous belt 151. The first stepper motor 14 is driven by one of the first synchronous pulleys 152. The first slider 17 is supported by the loading unit. The first slider 17 is fixedly connected to the first synchronous belt 151. The first slider 17 is slidably connected to the horizontal guide rail 16.
[0033] like Figure 7 As shown, in one embodiment, a cover assembly 18 is also included. The cover assembly 18 includes a high-temperature cloth 19 and a cover frame 20. The high-temperature cloth 19 covers the outside of the first synchronous belt 151 and moves with the first synchronous belt 151. The high-temperature cloth 19 is provided with a cover frame 20 that is the same size as the opening of each water bath. One end of the cover frame 20 is fixedly connected to the first slider 17. The cover assembly 18 can prevent water vapor from escaping and also has a heat preservation function.
[0034] like Figure 6As shown, in one embodiment, the vertical drive unit 21 includes a vertical guide rail 26, a second stepper motor 24, a second slider, a second synchronous belt 251, a second synchronous pulley 252, a counterweight 22, a guide rod 27, and a pulley block 23. The upper and lower ends of the vertical guide rail 26 are respectively provided with second synchronous pulleys 252. The two second synchronous pulleys 252 are connected by the second synchronous belt 251. The second stepper motor 24 is driven by one of the second synchronous pulleys 252. The second slider is connected to the loading unit. Two synchronous belts 251 are fixedly connected, and the second slider is slidably connected to the vertical guide rail 26. A guide rod 27 is connected between the upper and lower ends of the vertical guide rail 26. A counterweight 22 is mounted on the guide rod 27 and is fixedly connected to the second synchronous belt 251. When the second slider moves to the upper end of the vertical guide rail 26, the counterweight 22 is located at the lower end of the vertical guide rail 26. A pulley group 23 is set at the top of the vertical guide rail 26. The second slider and the counterweight 22 are connected by a suspension rope set on the pulley group 23. The vertical drive unit 21 is mainly used to realize the rising or falling of the water bath basket 4 in the water tank and the up and down vibration of the water bath basket 4 in the water tank. Since the water bath basket 4 is heavy when fully loaded, the component is designed with a counterweight 22, which is connected by the pulley group 23 to reduce the load requirements of the Z-axis motor.
[0035] like Figure 2 As shown, in one embodiment, a level gauge 31 is also included. The level gauge 31 is disposed on the outer wall of each water tank and is used to measure the water level inside the water tank.
[0036] like Figure 12 As shown, in one embodiment, the system further includes a frame assembly 35. The upper and lower parts of the frame assembly 35 are separated by a water bath frame. The water bath frame has openings corresponding to the top openings of each water bath. The thermal circulation unit 28 is located in the lower part of the frame assembly 35, while the horizontal drive unit 13 and vertical drive unit 21 are both located in the upper part. The frame assembly 35 primarily provides a platform for the installation and execution of each module. It mainly includes a profile frame 36, a screen assembly 37, an electrical box assembly 38, a sliding door assembly 39, and other auxiliary components. The electrical box assembly 38 is connected to the heating rod 48, the circulation pump 34, the first stepper motor 14, and the second stepper motor 24. The screen assembly 37 and the electrical box assembly 38 are both connected to the control unit.
[0037] like Figure 13 As shown, in one embodiment, this embodiment also provides a control method for the aforementioned fully automated high-throughput water bath PCR thermal cycler, comprising: Load the reaction plate into the loading unit; Obtain the PCR amplification experimental procedure set by the user; The user-defined PCR amplification experimental workflow is initiated. The PCR amplification experimental workflow execution process is as follows: The horizontal drive unit is controlled to move the loading unit directly above the target water bath tank; Determine whether the liquid level in each water bath has reached the upper liquid level. If not, replenish the liquid to the upper liquid level. If so, issue the target temperature for each water bath and control the heating components in each water bath to heat it. Once the temperature of each water tank reaches the starting temperature of the experiment, the remaining water tanks are subjected to timed temperature control, while the target water tank continues to be heated. When the real-time temperature of the target water bath reaches the target temperature, the vertical drive unit is controlled to lower the loading unit into the target water bath and fully immerse it in the water. Then, the loading unit is controlled to make small up-and-down swaying motions in the water, and the real-time temperature of the target water bath is maintained at the target temperature by controlling the heating and / or heat dissipation components. At the same time, the temperature of the next target water bath is controlled to be heated to the target temperature. Once the reaction in the target water bath is complete and the next target water bath reaches the target temperature, the vertical drive unit is controlled to lift the loading unit, and the horizontal drive unit is controlled to move the loading unit to the next target water bath, and the above experimental procedure is repeated.
[0038] The specific control process includes the following steps: Start; move the rack (i.e., water bath rack 12) to the interactive position (i.e., directly above the target water bath); the user loads the reaction basket (i.e., loads the water bath basket 4 carrying the reaction plate 9 onto the water bath rack 12); the user edits the experimental procedure on the screen component 37; start the experimental procedure; determine if the water level in the tank has reached the upper level; if not, replenish the target water tank to the upper level; if so, issue timed temperature control to each water tank through the control unit; determine if the water tank has reached the water tank temperature at the start of the experiment; if not, continue to control the temperature of the target water tank to achieve temperature control of each water tank, so that each water tank reaches the water tank temperature at the start of the experiment; if so, control the temperature of the target water bath to reach the required reaction temperature, and control the temperature of other water tanks in real time to maintain the water tank temperature at the start of the experiment; determine the temperature of the target water bath... If the target water bath temperature has not reached the required reaction temperature, wait for the target water bath temperature to be controlled to the temperature at the start of the experiment. If the waiting time is less than 90 seconds, repeat the above temperature control process from the start of the experiment to the required reaction temperature, and time the time it takes for the target water bath to reach the target temperature. Then determine if the experiment is complete. If the waiting time is not less than 90 seconds, control the target water bath temperature to the target temperature and time it. Then determine if the experiment is complete. If the experiment is not complete, issue an audible and visual reminder. If the experiment is complete, move the carrier (i.e., water bath basket hanger 12) to the standby position, and then determine if the user needs to perform the next round of heat preservation. If yes, repeat the above steps to control the target water temperature. If no, end the operation.
[0039] like Figure 8 As shown, the high-throughput water bath PCR thermal cycler of the present invention also includes a leakage protection switch 1, a power switch 2, and a device door 3. The device door 3 is an openable and closable device door. Both the leakage protection switch 1 and the power switch 2 are connected to the control unit. When using the high-throughput water bath PCR thermal cycler of the present invention, turn on the leakage protection switch 1, then turn on the power switch 2. The device will automatically power on, perform a self-test, and initialize. Place the water bath basket 4 into the device, edit the current experimental protocol via the touchscreen, close the device door 3, and start the experiment. The device will then perform PCR amplification according to the set protocol.
[0040] The specific working process of the device of this invention is as follows: For first-time use, pure water needs to be injected into the high-temperature water tank 6, low-temperature water tank 7, and ordinary water tank 8 through the water inlet 54 until the upper liquid level is reached. The water inlet 54 should be continuously connected to the pure water tank so that water can be automatically replenished during use. At the same time, an overflow outlet 5 is provided. When the water level in the tank exceeds the upper liquid level, the excess water is discharged through the overflow outlet 5 to prevent water from overflowing from the tank opening.
[0041] Place reaction plate 9 into reaction plate slot 10 of water bath basket 4. After placing all the reaction plates to be tested into water bath basket 4, install water bath basket baffle 11, place the water bath basket into the slide groove of water bath basket bracket 12 and push it backward. Figure 14 As shown, at this time, the positioning bead 42 in the slide groove is inserted into the positioning groove 43 of the water bath basket, and the front end of the water bath basket and the water bath basket bracket are flush.
[0042] Edit the PCR amplification protocol on the touchscreen, close the device door, and click "Start Process." The device will then begin the process. Wait for the process to complete.
[0043] The advantages of the high-throughput water bath PCR thermal cycler and its control method proposed in this invention compared with the prior art are as follows: This invention employs a detachable water bath basket structure. After the reaction plate to be tested is loaded into the water bath basket 4, the water bath basket 4 is then mounted onto the water bath basket rack 12 of the device for PCR water bath circulation. By using multiple of these water bath baskets in rotation, continuous and uninterrupted PCR water bath circulation of a large number of samples can be achieved, effectively increasing the device throughput. Furthermore, by configuring water bath baskets of different specifications, the device can support reaction plates of different sizes, such as 384-well deep plates, 384-well microplates, half-skirted 96-well PCR plates, and skirtless 96-well PCR plates, improving device compatibility and meeting diverse water bath PCR experimental needs.
[0044] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.
Claims
1. A fully automated high-throughput water bath PCR thermal cycler, characterized in that, include: The thermal circulation unit includes at least three water baths and a heat dissipation component. The top of each water bath is open. Each water bath includes a water tank, a heating component, a temperature detection module, and a water inlet. The heating component is located inside the water tank. The temperature detection module is used to detect the water temperature inside the water tank of each water bath. The heat dissipation component is used to dissipate heat and cool down at least one water bath. The water inlet is used to connect to an external water source. Loading unit; the loading unit is used for batch loading of reaction plates; A horizontal drive unit is used to drive the loading unit to move horizontally to directly above each water bath tank; The vertical drive unit is used to drive the loading unit to descend into the interior of each water bath tank, or to drive the loading unit to rise and leave the interior of each water bath tank. The control unit is connected to the heating components, horizontal drive units, vertical drive units, temperature detection modules, and heat dissipation components in each water bath.
2. The fully automated high-throughput water bath PCR thermal cycler according to claim 1, characterized in that, Each water bath is equipped with a return water inlet at the top. The heating components include heating pipes and heating rods. The heating pipes are horizontally installed at the bottom of the water tank, and the heating rods are installed inside the heating pipes. Several guide spray holes are installed on the top of the heating pipes, and a water inlet is installed at the bottom of the heating pipes. The water inlet and the return water inlet are connected by a circulation pump.
3. The fully automated high-throughput water bath PCR thermal cycler according to claim 2, characterized in that, The heat dissipation assembly includes a fan, heat pipes, and heat dissipation fins. The inlet end of the heat pipes is connected to the outlet of each circulation pump, and the outlet end of the heat pipes is connected to the inlet of the corresponding water bath. An inlet water temperature detection module is provided at the inlet. The fan is set close to the inside of the heat pipes, and the heat dissipation fins are set close to the outside of the heat pipes. The fan, the inlet water temperature detection module, and the control unit are connected.
4. The fully automated high-throughput water bath PCR thermal cycler according to claim 1, characterized in that, The horizontal drive unit includes a horizontal guide rail, a first stepper motor, a first slider, a first synchronous belt, and a first synchronous pulley. The front and rear ends of the horizontal guide rail are respectively provided with first synchronous pulleys. The two first synchronous pulleys are connected by the first synchronous belt. The first stepper motor is driven by one of the first synchronous pulleys. The first slider is supported by the loading unit. The first slider is fixedly connected to the first synchronous belt. The first slider is slidably connected to the horizontal guide rail.
5. The fully automated high-throughput water bath PCR thermal cycler according to claim 4, characterized in that, It also includes a cover assembly, which includes a high-temperature cloth and a cover frame. The high-temperature cloth covers the outside of the first synchronous belt and moves with the first synchronous belt. The high-temperature cloth is provided with a cover frame that is the same size as the opening of each water bath. One end of the cover frame is fixedly connected to the first slider.
6. The fully automated high-throughput water bath PCR thermal cycler according to claim 1, characterized in that, The vertical drive unit includes a vertical guide rail, a second stepper motor, a second slider, a second synchronous belt, a second synchronous pulley, a counterweight, a guide rod, and a pulley block. The upper and lower ends of the vertical guide rail are respectively equipped with second synchronous pulleys, which are connected by a second synchronous belt. The second stepper motor is driven by one of the second synchronous pulleys. The second slider is connected to the loading unit and is fixedly connected to the second synchronous belt. The second slider is slidably connected to the vertical guide rail. A guide rod connects the upper and lower ends of the vertical guide rail. The counterweight passes through the guide rod and is fixedly connected to the second synchronous belt. When the second slider moves to the upper end of the vertical guide rail, the counterweight is located at the lower end of the vertical guide rail. The pulley block is located at the top of the vertical guide rail, and the second slider and the counterweight are connected by a suspension rope mounted on the pulley block.
7. The fully automated high-throughput water bath PCR thermal cycler according to claim 5, characterized in that, The loading unit includes a water bath basket and a water bath basket bracket. The water bath basket bracket is fixedly connected to the second slider. The water bath basket bracket includes a spring, a water tank cover, and a bracket connected from top to bottom. The water tank cover fits into the top opening of each water bath. The upper end of the water tank cover is connected to the second slider through at least one spring. Horizontal guide grooves are provided on both sides of the bottom of the bracket. Horizontal slide rails that are adapted to and engaged with the guide grooves are provided on both sides of the top of the water bath basket.
8. The fully automated high-throughput water bath PCR thermal cycler according to claim 1, characterized in that, It also includes a level gauge, which is installed on the outer wall of each water tank and is used to measure the water level inside the tank.
9. The fully automated high-throughput water bath PCR thermal cycler according to claim 1, characterized in that, It also includes a frame assembly, the upper and lower parts of which are separated by a water bath frame. The water bath frame has openings corresponding to the top openings of each water bath. The heat circulation unit is located in the lower part of the frame assembly, while the horizontal drive unit and the vertical drive unit are located in the upper part of the frame assembly.
10. A control method for a fully automated high-throughput water bath PCR thermal cycler as described in any one of claims 1-9, characterized in that, include: Load the reaction plate into the loading unit; Obtain the PCR amplification experimental procedure set by the user; The user-defined PCR amplification experimental workflow is initiated. The PCR amplification experimental workflow execution process is as follows: The horizontal drive unit is controlled to move the loading unit directly above the target water bath tank; Determine whether the liquid level in each water bath has reached the upper liquid level. If not, replenish the liquid to the upper liquid level. If so, issue the target temperature for each water bath and control the heating components in each water bath to heat it. Once the temperature of each water tank reaches the starting temperature of the experiment, the remaining water tanks are subjected to timed temperature control, while the target water tank continues to be heated. When the real-time temperature of the target water bath reaches the target temperature, the vertical drive unit is controlled to lower the loading unit into the target water bath and fully immerse it in the water. Then, the loading unit is controlled to make small up-and-down swaying motions in the water. The heating and heat dissipation components are controlled to keep the real-time temperature of the target water bath at the target temperature. At the same time, the temperature of the next target water bath is controlled to be heated to the target temperature. Once the reaction in the target water bath is complete and the next target water bath reaches the target temperature, the vertical drive unit is controlled to raise the loading unit, and the horizontal drive unit is controlled to move the loading unit to the next target water bath, and the above experimental procedure is repeated.