An in vitro transcription apparatus for messenger ribonucleic acid

Abstract

The present disclosure provides an in vitro transcription device for messenger ribonucleic acid, which comprises a central controller, a consumable stack, a liquid workstation, a transfer module, a mechanical arm module and a reagent storage module, the central controller is used to control the operation of the liquid workstation and the mechanical arm module respectively, the consumable stack is used to store consumables involved in the in vitro transcription process, and the reagent storage module is used to store reagent containers involved in the in vitro transcription process. The present disclosure automatically carries consumables and reagent containers involved in the in vitro transcription process by the mechanical arm module, without human intervention, eliminates the cumulative position error in the carrying process of consumables and reagent containers through the transfer module, and realizes precise fully automated pipetting operation; at the same time, the plasmid PCR amplification link, the IVT reaction link, the DNaseI digestion link and the mRNA purification and enrichment link involved in the whole in vitro transcription process are all automatic operation, without human intervention, thereby improving the efficiency of mRNA in vitro transcription.

Description

Technical Field

[0001] This disclosure relates to the field of messenger ribonucleic acid (mRNA) preparation technology, and particularly to an in vitro transcription device for messenger ribonucleic acid. Background Technology

[0002] In the research and development and production of messenger RNA vaccines, in vitro transcription is the core step in mRNA synthesis. It requires four key steps to be completed in sequence: plasmid template PCR (Polymerase Chain Reaction) amplification, IVT (In Vitro Transcription) reaction (i.e., mRNA synthesis), DNase I (Deoxyribonuclease I) digestion (i.e., removal of residual deoxyribonuclease), and mRNA purification and enrichment (i.e., removal of impurities). The existing preparation process relies on manual operation. For example, pipetting (reagent transfer) requires manual use of pipettes and involves various reagents with different characteristics, such as enzymes, magnetic beads, and buffer solutions. Errors can easily occur due to differences in the force and speed of operation, making it impossible to achieve precise, fully automated pipetting. The purification process often uses the "column method," which requires manual transfer of samples to the chromatography column. This method is not compatible with automated equipment, and existing magnetic bead purification solutions have not optimized reagent dosage and temperature control parameters, resulting in low mRNA recovery rates and high levels of impurities (such as salts and proteins). Each step relies on dispersion instruments (PCR instruments, centrifuges, shakers, magnetic bead purifiers), which require manual sample transfer, resulting in low efficiency and a high risk of contamination. The problems of low efficiency, large human error, and poor batch reproducibility in traditional manual operations (such as column purification and reagent transfer) make it impossible to achieve precise, fully automated pipetting in the in vitro transcription process. Utility Model Content

[0003] The technical problem to be solved by this disclosure is to overcome the shortcomings of the existing technology in which the in vitro transcription process of messenger ribonucleic acid cannot achieve precise and fully automated pipetting operations and requires human intervention, and to provide an in vitro transcription device for messenger ribonucleic acid.

[0004] This disclosure solves the above-mentioned technical problems through the following technical solution:

[0005] In a first aspect, an in vitro transcription device for messenger ribonucleic acid is provided, the in vitro transcription device comprising a central controller, a consumable stack, a liquid workstation, a transfer module, a robotic arm module, and a reagent storage module;

[0006] The central controller is used to control the operation of the liquid workstation and the robotic arm module respectively;

[0007] The consumables stack is used to store consumables involved in the in vitro transcription process;

[0008] The reagent storage module is used to store reagent containers containing reagents involved in the in vitro transcription process;

[0009] The transfer module is used to receive the consumables and reagent containers transported by the robotic arm module;

[0010] The robotic arm module is used to move the consumables from the consumables stack to the first target position of the transfer module based on the first handling command sent by the central controller, and to move the consumables at the first target position to the liquid workstation;

[0011] The robotic arm module is used to move the reagent container from the reagent storage module to the second target position of the transfer module based on the second handling command sent by the central controller, and then move the reagent container at the second target position to the liquid workstation;

[0012] The liquid workstation is used to receive pipetting instructions sent by the central controller and to perform pipetting operations on the consumables and reagent containers based on the pipetting instructions.

[0013] Optionally, the transfer module is located between the consumables stack and the liquid workstation;

[0014] And / or, the transfer module is made of aluminum alloy;

[0015] And / or, the transfer module includes a top plate and a first side plate and a second side plate connected to and disposed opposite to the top plate, and a limiter is provided at the boundary of the top plate;

[0016] And / or, the top plate of the transfer module is provided with a target area for accommodating the consumables or reagent containers of matching size.

[0017] Optionally, the in vitro transcription device further includes a door opening device, the reagent storage module includes a refrigerator, and the reagents include low-temperature reagents;

[0018] The refrigerator is used to store the low-temperature reagent container containing the low-temperature reagent;

[0019] The door opening device is used to open the refrigerator door based on the door opening command sent by the central controller;

[0020] The door opening device is also used to close the refrigerator door based on a door closing command sent by the central controller.

[0021] Optionally, the door opening device includes a motor, a reduction mechanism, a connecting rod, a latch, and a door opening controller;

[0022] The buckle is connected to the door of the refrigerator, and the connecting rod, the reduction mechanism, and the motor are electrically connected in sequence.

[0023] The door opening controller is used to drive the motor to operate based on the door opening command or the door closing command sent by the central controller;

[0024] The motor is used to drive the reduction mechanism to perform a deceleration movement, thereby driving the connecting rod to move;

[0025] The connecting rod is used to drive the latch to open or close the refrigerator door.

[0026] Optionally, the in vitro transcription device further includes a sealing and tearing device, a shaking module, a centrifuge, and a PCR instrument;

[0027] The central controller is used to control the operation of the sealing and tearing device, the shaking module, the centrifuge, and the PCR instrument, respectively.

[0028] The consumables include an initial PCR plate and a first pipette tip box containing a first pipette tip.

[0029] The low-temperature reagent includes a plasmid template and a PCR reaction solution;

[0030] The liquid workstation is used to receive the initial PCR plate, the first pipette tip box, the plasmid template, and the PCR reaction solution container containing the PCR reaction solution transported by the robotic arm module, and to perform pipetting operations based on the first pipetting command sent by the central controller to obtain a PCR plate containing the amplification reaction solution.

[0031] The robotic arm module is also used to transport the PCR plate containing the amplification reaction solution to the sealing and tearing device via the transfer module for sealing operation, so as to obtain a sealed amplification PCR plate.

[0032] The robotic arm module is also used to transport the sealed amplification PCR plate to the oscillation module via the transfer module for oscillation to obtain an amplified and mixed PCR plate.

[0033] The robotic arm module is also used to transport the amplified and mixed PCR plate to the centrifuge via the transfer module for centrifugation to obtain the amplified centrifuged PCR plate.

[0034] The robotic arm module is also used to transport the amplified centrifuged PCR plate to the PCR instrument via the transfer module for PCR operation to obtain the plasmid amplification product plate.

[0035] Optionally, the in vitro transcription device further includes a heating module, and the low-temperature reagent further includes a pre-sealed IVT premix;

[0036] The robotic arm module is also used to transport the plasmid amplification product plate to the centrifuge via the transfer module for centrifugation to obtain a transcription centrifuged PCR plate.

[0037] The robotic arm module is also used to transport the transcription centrifuged PCR plate to the sealing and tearing device via the transfer module for tearing operation to obtain the transcription tearing PCR plate;

[0038] The robotic arm module is also used to transport the transcription membrane-tearing PCR plate to the liquid workstation via the transfer module;

[0039] The robotic arm module is also used to transport the container containing the IVT premixed liquid to the centrifuge via the transfer module for centrifugation, so as to centrifuge the IVT premixed liquid into centrifuged IVT premixed liquid; the robotic arm module is also used to transport the container containing the centrifuged IVT premixed liquid to the liquid workstation via the transfer module.

[0040] The robotic arm module is also used to transport the initial PCR plate in the consumable stack to the liquid workstation via the transfer module;

[0041] The liquid workstation is also used to receive the initial PCR plate, the first pipette tip box, and the container containing the centrifuged IVT premixed solution transported by the robotic arm module, and to perform pipetting operations based on the second pipetting command sent by the central controller to obtain a PCR plate containing IVT reaction solution.

[0042] The robotic arm module is also used to transport the PCR plate containing IVT reaction solution to the sealing and tearing device via the transfer module for sealing operation, so as to obtain a sealed PCR plate containing IVT reaction solution.

[0043] The robotic arm module is also used to transfer the sealed PCR plate containing IVT reaction solution to the shaking module via the transfer module for shaking operation, so as to obtain a mixed PCR plate containing IVT reaction solution.

[0044] The robotic arm module is also used to transfer the mixed PCR plate containing IVT reaction solution to the centrifuge via the transfer module for offline operation, so as to obtain a centrifuged IVT reaction solution PCR plate.

[0045] The robotic arm module is also used to transport the centrifuged IVT reaction solution PCR plate to the heating module via the transfer module;

[0046] The heating module is used to provide the corresponding first IVT reaction temperature to the centrifuged IVT reaction solution PCR plate based on the first IVT reaction temperature command sent by the central controller, so as to obtain the IVT product plate.

[0047] Optionally, the low-temperature reagent includes a pre-sealed PCR plate containing DNase I;

[0048] The robotic arm module is also used to transport the DNase I-containing PCR plate to the sealing and tearing device via the transfer module for tearing operation, so as to obtain a DNase I-containing tearing PCR plate.

[0049] The robotic arm module is also used to transport the DNase I-containing membrane-tear PCR plate to the liquid workstation via the transfer module.

[0050] The robotic arm module is also used to transport the IVT product plate to the centrifuge via the transfer module for centrifugation to obtain centrifuged IVT product plate;

[0051] The robotic arm module is also used to transport the centrifuged IVT product plate through the transfer module to the sealing and tearing machine for tearing operation to obtain the tearing IVT product plate.

[0052] The robotic arm module is also used to transport the film-tear IVT product plate to the liquid workstation via the transfer module.

[0053] The liquid workstation is used to receive the first pipette tip box, the membrane-tear IVT product plate and the membrane-tear PCR plate containing DNase I, and to perform pipetting operations based on the third pipetting command sent by the central controller to obtain a PCR plate containing DNase I digestion reaction solution.

[0054] The robotic arm module is also used to transport the PCR plate containing DNaseI digestion reaction solution to the sealing and tearing device via the transfer module for sealing operation, so as to obtain a sealed PCR plate containing DNaseI digestion reaction solution.

[0055] The robotic arm module is also used to transfer the sealed PCR plate containing DNaseI digestion reaction solution to the shaking module via the transfer module for shaking operation, so as to obtain a mixed PCR plate containing DNaseI digestion reaction solution.

[0056] The robotic arm module is also used to transfer the mixed PCR plate containing DNase I digestion reaction solution to the centrifuge via the transfer module for centrifugation to obtain a centrifuged PCR plate containing DNase I digestion reaction solution.

[0057] The robotic arm module is also used to transport the centrifuged PCR plate containing DNaseI digestion reaction solution to the heating module via the transfer module;

[0058] The heating module is used to provide the corresponding second IVT reaction temperature to the centrifuged PCR plate containing DNaseI digestion solution based on the second IVT reaction temperature command sent by the central controller, so as to obtain the enzyme digestion product plate.

[0059] Optionally, the robotic arm module is also used to transport the used DNase I-containing PCR plate to the sealing and tearing device via the transfer module for sealing operation;

[0060] And / or, the robotic arm module is also used to transport the first pipette tip box containing the first pipette tip, located at the liquid workstation, to the consumables stack via the transfer module;

[0061] And / or, the robotic arm module is also used to transport the enzyme digestion product plate to the refrigerator via the transfer module.

[0062] Optionally, the reagent storage module includes a buffer rack, and the reagents include room temperature reagents;

[0063] The buffer rack is used to store the room temperature reagent container containing the room temperature reagent;

[0064] The robotic arm module is used to transport the room-temperature reagent container to the liquid workstation via the transfer module;

[0065] Alternatively, the robotic arm module is used to transport the room-temperature reagent container to the buffer rack via the transfer module.

[0066] Optionally, the in vitro transcription device further includes a magnetic bead purification device;

[0067] The consumables include an initial deep-hole plate and a second pipette box equipped with a second pipette tip, wherein the pipetting capacity range of the second pipette tip is greater than that of the first pipette tip;

[0068] The room-temperature reagent includes magnetic beads, binding buffer, washing buffer, and elution buffer;

[0069] The robotic arm module is also used to transport the first liquid storage tank containing the magnetic beads, the second liquid storage tank containing the binding buffer, the third liquid storage tank containing the washing buffer, and the fourth liquid storage tank containing the elution buffer from the buffer rack to the liquid workstation via the transfer module, respectively.

[0070] The robotic arm module is also used to transport the enzyme digestion product plate to the centrifuge via the transfer module for centrifugation to obtain the centrifuged enzyme digestion product plate.

[0071] The robotic arm module is also used to transport the centrifuged enzyme digestion product plate to the sealing and tearing device via the transfer module for tearing operation to obtain the tearing enzyme digestion product plate.

[0072] The robotic arm module is also used to transport the membrane-peeling enzyme digestion product plate and the initial deep well plate to the liquid workstation via the transfer module;

[0073] The liquid workstation is also used to receive the initial deep well plate, the first pipette tip box, the second pipette tip box, the first liquid storage tank, the second liquid storage tank, the third liquid storage tank, the fourth liquid storage tank and the membrane-tear enzyme digestion product plate transported by the robotic arm module, and to perform pipetting operations based on the fourth pipetting command sent by the central controller to obtain a deep well plate for purification and enrichment of reagents.

[0074] The robotic arm module is also used to transport the purification and enrichment reagent deep well plate to the magnetic bead purifier via the transfer module.

[0075] The magnetic bead purifier is used to purify the purification enrichment reagent deep-well plate based on the purification instructions sent by the central controller, so as to obtain the mRNA deep-well plate.

[0076] Optionally, the robotic arm module is also used to transport the cleaned magnetic bead deep well plate after the purification operation to the magnetic bead purifier via the transfer module;

[0077] The liquid workstation is also used to receive the first suction tip box, the second suction tip box, the first liquid storage tank, the second liquid storage tank, the third liquid storage tank, the fourth liquid storage tank and the cleaning magnetic bead deep well plate transported by the robotic arm module, and to perform a liquid transfer operation based on the fifth liquid transfer command sent by the central controller to obtain the magnetic bead recovery reagent deep well plate.

[0078] The robotic arm module is also used to transport the magnetic bead recovery reagent deep well plate to the magnetic bead purifier via the transfer module;

[0079] The magnetic bead purifier is used to perform a recovery operation on the cleaned magnetic beads in the magnetic bead recovery reagent deep well plate based on the recovery command sent by the central controller, so as to obtain a recovered magnetic bead deep well plate containing the recovered magnetic beads.

[0080] Optionally, the robotic arm module is also used to transport the mRNA deep well plate or the recycled magnetic bead deep well plate to the buffer rack via the transfer module;

[0081] And / or, the second pipette tip has a pipetting capacity range of 300 μL, and the first pipette tip has a pipetting capacity range of 20 μL.

[0082] Based on common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of this disclosure.

[0083] The positive and progressive effects of this disclosure are as follows:

[0084] The messenger RNA in vitro transcription device disclosed herein includes a central controller, a consumable stack, a liquid workstation, a transfer module, a robotic arm module, and a reagent storage module. The central controller controls the operation of the liquid workstation and the robotic arm module respectively. The consumable stack stores consumables involved in the in vitro transcription process, and the reagent storage module stores reagents involved in the in vitro transcription process. The robotic arm module automatically transports consumables and reagents involved in the in vitro transcription process without human intervention. The transfer module eliminates the cumulative positional error (deviation) during the handling of consumables and reagents, solving the problem of continuous accumulation of positional error during movement and the occurrence of misplacement in the designated position of the liquid workstation during transfer. This achieves precise, fully automated pipetting operations, resulting in good batch reproducibility of messenger RNA in vitro transcription and improving the efficiency of messenger RNA in vitro transcription. Attached Figure Description

[0085] Figure 1 A first structural schematic diagram of the in vitro transcription device for messenger ribonucleic acid provided in this disclosure;

[0086] Figure 2 This is a first structural schematic diagram of the transfer module in the in vitro transcription device for messenger ribonucleic acid provided in this disclosure;

[0087] Figure 3 This is a second structural schematic diagram of the transfer module in the in vitro transcription device for messenger ribonucleic acid provided in this disclosure;

[0088] Figure 4 This is a schematic diagram of the third structure of the transfer module in the in vitro transcription device for messenger ribonucleic acid provided in this disclosure;

[0089] Figure 5 This is a second structural schematic diagram of the in vitro transcription device for messenger ribonucleic acid provided in this disclosure;

[0090] Figure 6 This is a first structural schematic diagram of the opening device in the in vitro transcription apparatus for messenger ribonucleic acid provided in this disclosure;

[0091] Figure 7 This is a second structural schematic diagram of the opening device in the in vitro transcription apparatus for messenger ribonucleic acid provided in this disclosure;

[0092] Figure 8 This is a third structural schematic diagram of the opening device in the in vitro transcription apparatus for messenger ribonucleic acid provided in this disclosure. Detailed Implementation

[0093] The present disclosure is further illustrated below by way of embodiments, but the present disclosure is not limited to the scope of the embodiments described herein.

[0094] The prefixes such as "first" and "second" used in this disclosure are merely for distinguishing different descriptive objects and do not limit the position, order, priority, quantity, or content of the described objects. The use of ordinal numbers and other prefixes used to distinguish descriptive objects in this disclosure does not constitute a limitation on the described objects. The description of the described objects is given in the claims or the context of the embodiments, and should not be construed as an unnecessary limitation. Furthermore, in the description of this embodiment, unless otherwise stated, "multiple" means two or more.

[0095] This embodiment provides an in vitro transcription device for messenger ribonucleic acid, such as... Figure 1 As shown, the in vitro transcription device includes a central controller 1, a consumable stack 2, a liquid workstation 3, a transfer module 4, a robotic arm module 5, and a reagent storage module 6;

[0096] The central controller 1 is used to control the operation of the liquid workstation 3 and the robotic arm module 5 respectively;

[0097] Consumables stack 2 is used to store consumables involved in the in vitro transcription process;

[0098] The reagent storage module 5 is used to store reagent containers containing reagents involved in the in vitro transcription process;

[0099] Transfer module 4 is used to receive consumables and reagent containers transported by robotic arm module 5;

[0100] The robotic arm module 5 is used to move consumables from the consumable stack 2 to the first target position of the transfer module 4 based on the first handling command sent by the central controller 1, and to move the consumables at the first target position to the liquid workstation 3;

[0101] The robotic arm module 5 is used to move reagent containers from the reagent storage module 6 to the second target position of the transfer module 4 based on the second handling command sent by the central controller, and then move the reagent containers at the second target position to the liquid workstation 3;

[0102] The liquid workstation 3 is used to receive pipetting instructions sent by the central controller 1 and to perform pipetting operations on consumables and reagent containers based on the pipetting instructions.

[0103] The central controller is compatible with various software to achieve fully automated control of various instruments and equipment (such as liquid workstations and robotic arm modules), and supports the export and traceability of operation logs.

[0104] The consumables stack is used to store consumables involved in the in vitro transcription process, such as unused and used PCR plates, deep well plates, and other experimental consumables. It supports automatic identification of plate position status, automatic grabbing of consumables by the robotic arm module, and software control of the stack position and recording of consumable information by the central control module, thereby realizing automated grabbing and movement of consumables.

[0105] The robotic arm module includes a robotic arm and grippers. Under the control of a central controller, it can automatically transport and transfer consumables and reagents between modules (devices), and has high-precision positioning and gripping capabilities. The robotic arm module disclosed herein is an existing mature device. This disclosure does not limit the specific structure of the robotic arm module, as long as it can achieve the corresponding functions.

[0106] The liquid workstation, under the control of a central controller, can perform high-precision liquid handling operations. The liquid workstation disclosed herein is a mature, existing device. This disclosure does not limit the specific structure of the liquid workstation, as long as it can achieve the corresponding functions.

[0107] The transfer module is used to receive consumables and reagent containers transported by the robotic arm module. When the robotic arm module transports consumables and reagent containers, it needs to use the transfer module as a bridge for transfer, so that a certain consumable is located at the first target position of the transfer module and a certain reagent container is located at the second target position of the transfer module. This eliminates the cumulative positional error during the transportation of consumables and reagent containers, and solves the problem of positional error accumulating during the movement process, which may result in the container not being placed in the designated position of the liquid workstation during transfer.

[0108] The robotic arm can move in the horizontal, vertical, and depth (or forward / backward) directions, corresponding to a three-dimensional coordinate system. Therefore, the cumulative error in the horizontal, vertical, and depth (or forward / backward) directions can be reduced by using a transfer module.

[0109] The messenger RNA in vitro transcription device of this embodiment uses a robotic arm module to automatically transport consumables and reagent containers involved in the in vitro transcription process without human intervention. The transfer module eliminates the cumulative positional error during the transportation of consumables and reagent containers, solving the problem of continuous accumulation of positional errors during movement and the occurrence of misplacement in the designated position of the liquid workstation during transport. It achieves precise and fully automated pipetting operation, and the batch reproducibility of messenger RNA in vitro transcription is good, thus improving the efficiency of messenger RNA in vitro transcription.

[0110] In an alternative implementation, the transfer module is located between the consumables stack and the liquid workstation.

[0111] The transfer module is located between the consumables stack and the liquid workstation, which reduces the operating distance of the robotic arm module and allows for a more compact layout of the messenger ribonucleic acid in vitro transcription device, reducing the area occupied by the messenger ribonucleic acid in vitro transcription device.

[0112] The transfer module can also be located at the center of the entire device to achieve the requirement of a shorter total path.

[0113] In one alternative embodiment, the transfer module is made of aluminum alloy.

[0114] Specifically, the aluminum alloy can be 6063 aluminum alloy, which is a wrought aluminum alloy with magnesium and silicon as the main alloying elements, with a magnesium content of 0.45%-0.9% and a silicon content of 0.2%-0.6%.

[0115] In an alternative implementation, such as Figure 2 , Figure 3 and Figure 4 As shown, the transfer module 4 includes a top plate 41 and a first side plate 42 and a second side plate 43 connected to and disposed opposite to the top plate 41. A limiter 44 is provided at the boundary of the top plate 41.

[0116] The first and second side plates are used to support the top plate, and the first and second side plates are connected and fixed to the top plate by screws.

[0117] In an alternative embodiment, the transfer module further includes a height adjustment unit for adjusting the height of the top plate of the transfer module relative to the consumable stack.

[0118] The top plate is used to place consumables and reagents transported by the robotic arm module. The consumable stack is a multi-layered device with several holes. The height of the top plate relative to the consumable stack is adjusted by the height adjustment unit so that the top plate of the transfer module is adapted to the position of the corresponding consumable. Only the robotic arm module needs to move horizontally, without vertical (up and down) movement, which further ensures the transport accuracy and reduces the cumulative error in the transport process.

[0119] In an alternative embodiment, the top plate of the transfer module is provided with a target area for accommodating consumable or reagent containers of matching size.

[0120] The size of the target area is set with reference to the base size of various consumables or reagent containers involved in the transcription device (all consumables are standard consumables, all reagent containers are standard containers, and the base size is consistent). For example, the size of the target area is obtained by adding 2-3mm to the length and width of the consumable base size.

[0121] By setting a target area at a fixed location, the robotic arm can accurately place consumables or reagent containers of matching size on the top plate, further reducing the cumulative error in the handling process.

[0122] In an alternative implementation, such as Figure 5As shown, the in vitro transcription device also includes a door opening device 7, and the reagent storage module 6 includes a refrigerator 61, and the reagents include low-temperature reagents.

[0123] Refrigerator 61 is used to store containers containing cryogenic reagents;

[0124] The door opening device 7 is used to open the door of the refrigerator 61 based on the door opening command sent by the central controller 1;

[0125] The door opening device 7 is also used to close the door of the refrigerator 61 based on the door closing command sent by the central controller 1.

[0126] Refrigerators can store various low-temperature reagents involved in the in vitro transcription process.

[0127] Under the control of the central controller, the door opening device can automatically open or close the refrigerator door. The robotic arm module is used to transport low-temperature reagent containers to the refrigerator via the transfer module, or to transport low-temperature reagent containers stored in the refrigerator to the liquid workstation via the transfer module, thus realizing the automated handling of low-temperature reagents in the in vitro transcription device.

[0128] In an alternative implementation, such as Figure 6 , Figure 7 and Figure 8 As shown, the door opening device 7 includes a motor 71, a reduction mechanism 72, a connecting rod 73, a latch 74, and a door opening controller (not shown in the figure).

[0129] The buckle 74 is connected to the door 611 of the refrigerator 61, and the connecting rod 73, the reduction mechanism 72, and the motor 71 are electrically connected in sequence.

[0130] The door opening controller is used to drive the motor 71 to operate based on the door opening or closing command sent by the central controller 1;

[0131] The motor 71 is used to drive the reduction mechanism 72 to perform a deceleration motion so as to drive the connecting rod 73 to move;

[0132] The connecting rod 73 is used to drive the latch 74 to open or close the door 611 of the refrigerator 61.

[0133] Under the control of the central control module, the door opening device can automatically open or close the refrigerator door, transforming an ordinary refrigerator into one that can automatically open and close the door. Moreover, the device has a simple structure, can be adapted to different types of refrigerators, and has a wide range of applications.

[0134] When it is necessary to open or close the door, the central control module sends an opening command or closing command to the door opening controller. The door opening controller controls the motor to move according to the command until the motor drives the reduction mechanism to move, which in turn drives the linkage to move. The linkage drives the latch to open or close the refrigerator door.

[0135] Specifically, a position sensor can be installed in the door opening device to detect the position of the door, thereby precisely controlling the opening angle of the refrigerator door.

[0136] In an alternative implementation, such as Figure 5 As shown, the in vitro transcription device also includes a sealing and tearing device 8, a shaking module 9, a centrifuge 10, and a PCR instrument 11;

[0137] The central controller 1 is used to control the operation of the sealing and tearing device 8, the shaking module 9, the centrifuge 10 and the PCR instrument 11 respectively;

[0138] Consumables include the initial PCR plate and the first pipette tip box containing the first pipette tip;

[0139] The low-temperature reagents include plasmid templates and PCR reaction solutions;

[0140] The liquid workstation 3 is used to receive the initial PCR plate, the first pipette tip box, the plasmid template and the PCR reaction solution container containing the PCR reaction solution transported by the robotic arm module, and to perform pipetting operations based on the first pipetting command sent by the central controller to obtain a PCR plate containing the amplification reaction solution.

[0141] The robotic arm module 5 is also used to transport the PCR plate containing the amplification reaction solution to the sealing and tearing device 8 via the transfer module 4 for sealing operation, so as to obtain the sealed amplification PCR plate.

[0142] The robotic arm module 5 is also used to transport the sealed amplification PCR plate to the shaking module 9 via the transfer module 4 for shaking to obtain the amplified and mixed PCR plate.

[0143] The robotic arm module 5 is also used to transport the amplified and mixed PCR plate to the centrifuge 10 via the transfer module 4 for centrifugation to obtain the amplified centrifuged PCR plate.

[0144] The robotic arm module 5 is also used to transport the amplified centrifuged PCR plate to the PCR instrument 11 via the transfer module 4 for PCR operation to obtain the plasmid amplification product plate.

[0145] Specifically, the initial PCR plate is an empty, unused PCR plate.

[0146] The sealing and tearing device, shaking module, centrifuge, and PCR instrument in this embodiment are all existing and mature devices. This disclosure does not limit the structure of the sealing and tearing device, shaking module, centrifuge, and PCR instrument, as long as they can achieve the corresponding functions.

[0147] Users can configure the operating programs of the sealing and tearing device, shaking module, centrifuge, and PCR instrument separately through the software, thereby enabling the central controller to control the operation of the sealing and tearing device, shaking module, centrifuge, and PCR instrument.

[0148] For example, users can set PCR operation (PCR reaction) conditions (including initial denaturation temperature, initial denaturation temperature duration, denaturation temperature within cycles, denaturation duration within cycles, annealing temperature, annealing duration, extension temperature within cycles, extension duration within cycles, number of cycles, extension temperature, and extension time) and the number of sample columns. The PCR instrument automatically matches the corresponding process and supports one-click start, and will automatically alarm in case of abnormality.

[0149] The in vitro transcription apparatus of this embodiment automates the plasmid PCR amplification step in the in vitro transcription process, eliminating the need for manual intervention and providing a plasmid amplification product plate, thus laying the foundation for subsequent in vitro transcription (IVT reaction) steps.

[0150] In an optional implementation, the robotic arm module is also used to transport the used plasmid template and used PCR reaction solution via a transfer module to a sealing and tearing device for sealing. This prevents the used plasmid template and used PCR reaction solution from being exposed to the air and causing air pollution.

[0151] In an optional implementation, the robotic arm module is also used to transport the first pipette tip located in the liquid workstation to the transfer module in the plasmid PCR amplification process, and to transport the first pipette tip located in the transfer module to the consumable stack to facilitate the use of the first pipette tip in subsequent processes.

[0152] In an alternative implementation, such as Figure 5 As shown, the in vitro transcription device also includes a heating module 12, and the low-temperature reagent also includes a pre-sealed IVT premix;

[0153] The robotic arm module 5 is also used to transport the plasmid amplification product plate to the centrifuge 10 via the transfer module 4 for centrifugation to obtain the transcription centrifuged PCR plate.

[0154] The robotic arm module 5 is also used to transport the transcription centrifuged PCR plate to the sealing and tearing device 8 via the transfer module 4 for the tearing operation to obtain the transcription tearing PCR plate.

[0155] Robotic arm module 5 is also used to transport transcription membrane-tearing PCR plates to liquid workstation 3 via transfer module 4;

[0156] The robotic arm module 5 is also used to transport the container containing the IVT premixed liquid to the centrifuge 10 via the transfer module 4 for centrifugation to centrifuge the IVT premixed liquid into centrifuged IVT premixed liquid.

[0157] The robotic arm module 5 is also used to transport containers containing centrifuged IVT premixed liquid to the liquid workstation 3 via the transfer module 4;

[0158] The robotic arm module 5 is also used to transport the initial PCR plates in the consumable stack 2 to the liquid workstation 3 via the transfer module 4;

[0159] The liquid workstation 3 is also used to receive the initial PCR plate, the first pipette tip box and the container containing the centrifuged IVT premixed solution transported by the robotic arm module 5, and to perform pipetting operations based on the second pipetting command sent by the central controller 1 to obtain a PCR plate containing IVT reaction solution.

[0160] The robotic arm module 5 is also used to transport the PCR plate containing IVT reaction solution to the sealing and tearing device 8 via the transfer module 4 for sealing operation, so as to obtain a sealed PCR plate containing IVT reaction solution.

[0161] The robotic arm module 5 is also used to transfer the sealed PCR plate containing IVT reaction solution to the shaking module 9 via the transfer module 4 for shaking operation, so as to obtain a mixed PCR plate containing IVT reaction solution.

[0162] The robotic arm module 5 is also used to transfer the mixed PCR plate containing IVT reaction solution to the centrifuge 10 via the transfer module 4 for offline operation, so as to obtain the centrifuged IVT reaction solution PCR plate.

[0163] Robotic arm module 5 is also used to transport centrifuged IVT reaction solution PCR plates to heating module 12 via transfer module 4;

[0164] The heating module 12 is used to provide the corresponding first IVT reaction temperature to the centrifuged IVT reaction solution PCR plate based on the first IVT reaction temperature command sent by the central controller 1, so as to obtain the IVT product plate.

[0165] The in vitro transcription device of this embodiment automates the IVT reaction process during in vitro transcription, eliminating the need for manual intervention and allowing the production of IVT product plates, thus laying the foundation for the subsequent DNaseI digestion process.

[0166] In an optional embodiment, the robotic arm module 5 is also used to transport the used IVT premix and / or the used plasmid amplification product plate to the sealing and tearing device via the transfer module for sealing operation, so as to prevent the used plasmid amplification product plate and / or IVT premix from being exposed to the air and causing air pollution.

[0167] In an optional embodiment, the robotic arm module 5 is also used to transport the first pipette tip located in the liquid workstation in the IVT reaction process to the consumable stack via a transfer module, so as to facilitate the use of the first pipette tip in subsequent processes.

[0168] In an optional embodiment, the low-temperature reagent includes a pre-sealed PCR plate containing DNase I;

[0169] The robotic arm module 5 is also used to transport the PCR plate containing DNase I to the sealing and tearing device 8 via the transfer module 4 for the tearing operation, so as to obtain the PCR plate containing DNase I.

[0170] Robotic arm module 5 is also used to transport DNase I-containing peelable PCR plates to liquid workstation 3 via transfer module 4;

[0171] The robotic arm module 5 is also used to transport the IVT product plate to the centrifuge 10 via the transfer module 4 for centrifugation to obtain centrifuged IVT product plate.

[0172] The robotic arm module 5 is also used to transport the centrifuged IVT product plate through the transfer module 4 to the sealing and tearing machine 8 for the tearing operation to obtain the tearing IVT product plate.

[0173] The robotic arm module 5 is also used to transport the film-tear IVT product plate to the liquid workstation 3 via the transfer module 4;

[0174] The liquid workstation 3 is used to receive the first pipette tip box, the membrane-torn IVT product plate and the membrane-torn PCR plate containing DNase I, and to perform pipetting operations based on the third pipetting command sent by the central controller to obtain a PCR plate containing DNase I digestion reaction solution.

[0175] The robotic arm module 5 is also used to transport the PCR plate containing DNase I digestion reaction solution to the sealing and tearing device 8 via the transfer module 4 for sealing operation, so as to obtain a sealed PCR plate containing DNase I digestion reaction solution.

[0176] The robotic arm module 5 is also used to transfer the sealed PCR plate containing DNase I digestion reaction solution to the shaking module 9 via the transfer module 4 for shaking operation, so as to obtain a mixed PCR plate containing DNase I digestion reaction solution.

[0177] The robotic arm module 5 is also used to transfer the mixed PCR plate containing DNaseI digestion reaction solution to the centrifuge 10 via the transfer module for centrifugation to obtain a centrifuged PCR plate containing DNaseI digestion reaction solution.

[0178] The robotic arm module 5 is also used to transfer the centrifuged PCR plate containing DNaseI digestion reaction solution to the heating module 12 via the transfer module 4;

[0179] The heating module 12 is used to provide the corresponding second IVT reaction temperature to the centrifuged PCR plate containing DNaseI digestion reaction solution based on the second IVT reaction temperature command sent by the central controller 1, so as to obtain the enzyme digestion product plate.

[0180] The in vitro transcription device of this embodiment automates the DNaseI digestion process during in vitro transcription without human intervention, and can obtain enzyme digestion product plates, laying the foundation for subsequent mRNA purification and enrichment and magnetic bead recovery.

[0181] In an optional implementation, the robotic arm module is also used to transport the used DNase I-containing peelable PCR plate to the sealing peeler via a transfer module for sealing operation; to prevent the used DNase I-containing peelable PCR plate from being exposed to the air and causing air pollution.

[0182] In an optional implementation, the robotic arm module is also used to transport the first pipette tip box containing the first pipette tip, located in the liquid workstation during the DNaseI digestion process, to the consumables stack via a transfer module, so as to facilitate the use of the first pipette tip in subsequent processes.

[0183] In an optional implementation, the robotic arm module is also used to transport the enzyme digestion product plate to a refrigerator via a transfer module.

[0184] The enzyme digestion product plates need to be stored at low temperatures, therefore, the enzyme digestion product plates need to be moved to a refrigerator.

[0185] In an alternative implementation, such as Figure 5 As shown, the reagent storage module 6 includes a buffer rack 62, and the reagents include room temperature reagents;

[0186] The buffer rack 62 is used to store containers containing room temperature reagents;

[0187] Robotic arm module 5 is used to transport room temperature reagent containers to liquid workstation 3 via transfer module 4;

[0188] Alternatively, the robotic arm module 5 is used to transport room-temperature reagent containers to the buffer rack via the transfer module 4.

[0189] The in vitro transcription device of this embodiment stores room-temperature reagents and intermediate products involved in the in vitro transcription process through a buffer rack, and realizes automated handling of room-temperature reagents or intermediate products through a robotic arm module.

[0190] In an alternative implementation, such as Figure 5 As shown, the in vitro transcription device also includes a magnetic bead purification device 13;

[0191] The consumables include an initial deep-well plate and a second pipette box equipped with a second pipette tip, the second pipette tip having a pipetting capacity range greater than that of the first pipette tip;

[0192] The room temperature reagents include magnetic beads, binding buffer, washing buffer, and elution buffer;

[0193] The robotic arm module 5 is also used to transfer the first liquid storage tank containing magnetic beads, the second liquid storage tank containing binding buffer, the third liquid storage tank containing washing buffer, and the fourth liquid storage tank containing elution buffer on the buffer rack 62 to the liquid workstation 3 via the transfer module 4, respectively.

[0194] The robotic arm module 5 is also used to transport the enzyme digestion product plate to the centrifuge via the transfer module 4 for centrifugation to obtain the centrifuged enzyme digestion product plate.

[0195] The robotic arm module 5 is also used to transport the centrifuged enzyme digestion product plate to the sealing and tearing device 8 via the transfer module 4 for tearing operation to obtain the tearing enzyme digestion product plate.

[0196] Robotic arm module 5 is also used to transport the membrane-tear enzyme digestion product plate and the initial deep well plate to the liquid workstation 3 via the transfer module;

[0197] The liquid workstation is also used to receive the initial deep well plate, the first pipette tip box, the second pipette tip box, the first liquid storage tank, the second liquid storage tank, the third liquid storage tank, the fourth liquid storage tank, and the membrane-torn enzyme digestion product plate transported by the robotic arm module, and to perform pipetting operations based on the fourth pipetting command sent by the central controller to obtain the purification and enrichment reagent deep well plate.

[0198] The robotic arm module 5 is also used to transport the purification and enrichment reagent deep well plate to the magnetic bead purifier 13 via the transfer module 4;

[0199] The magnetic bead purifier 13 is used to purify deep-well plates containing purification enrichment reagents based on purification instructions sent by the central controller, in order to obtain mRNA deep-well plates.

[0200] Specifically, the second pipette tip has a pipetting capacity range of 300 μL (large volume buffer), and the first pipette tip has a pipetting capacity range of 20 μL (micro reagent). The liquid workstation performs pipetting operations according to the fourth pipetting instruction to obtain a purified and enriched reagent deep-well plate. The liquid workstation calibrates pipetting accuracy through software, controlling the CV (coefficient of variation) value to ≤5%, and supports "multiple aspiration, single dispensing" and "single aspiration, multiple dispensing" modes, such as evenly distributing 100 μL magnetic beads into 12 wells, reducing the number of operation steps.

[0201] Specifically, the correspondence and experimental functions of the magnetic bead purification instrument are as follows:

[0202] Position 1: Denatured at 65℃ (nucleic acid denaturation) and bound to magnetic beads;

[0203] Position 2: Binding Buffer cleans the magnetic beads (removes impurities);

[0204] Position 3: Binding Buffer cleans the magnetic beads (removes impurities);

[0205] Position 4: Washing Buffer cleans non-specifically bound nucleic acids;

[0206] Position 5: Washing Buffer cleans non-specifically bound nucleic acids.

[0207] Position 6: Elution with 75℃ Elution Buffer (collect mRNA);

[0208] Position 7: Washing Buffer cleans used magnetic beads and places used magnetic rod sleeves.

[0209] Position 8: Place the magnetic rod sleeve to prevent contamination between samples.

[0210] Specifically, reagent dosages were optimized as follows: magnetic beads (50 μL / well), binding buffer (200 μL / well), washing buffer (150 μL / well), and elution buffer (100 μL / well). Temperature coordination was implemented: nucleic acid denaturation at 65℃ (destroying non-specific binding), and nucleic acid elution at 75℃ (improving mRNA recovery), with a temperature accuracy of ±0.5℃.

[0211] After the robotic arm module transports the purification enrichment reagent deep-well plate to the magnetic bead purifier, the magnetic bead purifier automatically runs the purification operation under the control of the central controller. After the operation is completed, the deep-well plate containing the final product (i.e., mRNA) in position 6 will be transferred to the transfer module via the robotic arm module, and then transferred from the transfer module to the buffer rack. The deep-well plates in positions 1, 2, 3, 4, 5, and 8 of the magnetic bead purifier will be transferred to the transfer module via the robotic arm module, and then transferred from the transfer module to the consumables stack.

[0212] The in vitro transcription device in this embodiment automates the mRNA purification and enrichment process during in vitro transcription. This automates all steps involved in the entire in vitro transcription process, including plasmid PCR amplification, IVT reaction, DNase I digestion, and mRNA purification and enrichment, eliminating the need for manual intervention and improving the efficiency of in vitro mRNA transcription.

[0213] In an optional implementation, the robotic arm module is also used to transport the cleaned magnetic bead deep-well plate after purification to the magnetic bead purifier via a transfer module.

[0214] The liquid workstation is also used to receive the first pipette tip box, the second pipette tip box, the first liquid storage tank, the second liquid storage tank, the third liquid storage tank, the fourth liquid storage tank and the cleaning magnetic bead deep well plate transported by the robotic arm module, and to perform pipetting operations based on the fifth pipetting command sent by the central controller to obtain the magnetic bead recovery reagent deep well plate.

[0215] The robotic arm module is also used to transfer magnetic bead recovery reagent deep well plates to the magnetic bead purifier via the transfer module;

[0216] The magnetic bead purifier is used to recover cleaned magnetic beads in a deep-well plate containing recovered magnetic beads based on recovery instructions sent by a central controller.

[0217] Specifically, after obtaining the magnetic bead recovery reagent deep well plate, the robotic arm module is also used to transport the magnetic bead recovery reagent deep well plate to the buffer rack via the transfer module. When magnetic bead recovery is required, the robotic arm module transports the magnetic bead recovery reagent deep well plate on the buffer rack to the magnetic bead purifier via the transfer module.

[0218] During the purification process, position 7 of the magnetic bead purifier functions to wash the used magnetic beads with washing buffer and place the used magnetic rod sleeve. The product obtained at position 7 is a purified magnetic bead deep-well plate. The robotic arm module transports the cleaned magnetic bead deep-well plate to position 9 of the magnetic bead purifier via a transfer module. The robotic arm module then retrieves the magnetic bead recovery reagent deep-well plate and transports it back to the magnetic bead purifier via the transfer module. The specific correspondence and experimental functions are as follows:

[0219] Position 9: Washing Buffer cleans used magnetic beads and places used magnetic rod sleeves.

[0220] Position 10: NaOH removes residual nucleic acid from used magnetic beads;

[0221] Position 11: Elution Buffer cleans the magnetic beads (removes impurities);

[0222] Position 12: Binding Buffer cleans the magnetic beads (removes impurities) and provides a buffer environment;

[0223] Position 13: Binding Buffer provides an environment for long-term storage of the magnetic bead;

[0224] After the robotic arm module transports the magnetic bead recovery reagent deep well plate to the magnetic bead purifier, the magnetic bead purifier automatically runs under the control of the central controller to perform the recovery operation. After the operation is completed, the robotic arm module transports the deep well plate containing the recovered magnetic beads in position 13 to the buffer rack via the transfer module. The deep well plates in positions 9-12 of the magnetic bead purifier are transferred to the transfer module via the robotic arm module, and then transferred from the transfer module to the consumable stack.

[0225] After the entire process is completed, the deep well plate containing the final product (mRNA) and the deep well plate with the recovered magnetic beads (which can be reused 5 times) are stored in the buffer rack.

[0226] The in vitro transcription device in this embodiment automates the magnetic bead retrieval process during in vitro transcription, eliminating the need for manual intervention to obtain a deep-well plate with retrieved magnetic beads. The retrieved magnetic beads in the deep-well plate can be reused approximately 5 times, reducing consumable costs.

[0227] In an alternative implementation, such as Figure 5 As shown, the in vitro transcription device also includes an alarm light 14, which automatically triggers an audible and visual alarm when the in vitro transcription device malfunctions, facilitating timely intervention and handling.

[0228] In an alternative embodiment, the robotic arm module of the in vitro transcription device includes a rotating unit for rotating the robotic arm so that the robotic arm rotates the consumables or reagents in the grippers to place the consumables or reagents in a preset orientation.

[0229] While specific embodiments of this disclosure have been described above, those skilled in the art should understand that these are merely illustrative examples, and the scope of protection of this disclosure is defined by the appended claims. Those skilled in the art can make various changes or modifications to these embodiments without departing from the principles and essence of this disclosure, but all such changes and modifications fall within the scope of protection of this disclosure.

Claims

1. An in vitro transcription device for messenger ribonucleic acid, characterized in that, The in vitro transcription device includes a central controller, a consumables stack, a liquid workstation, a transfer module, a robotic arm module, and a reagent storage module; The central controller is used to control the operation of the liquid workstation and the robotic arm module respectively; The consumables stack is used to store consumables involved in the in vitro transcription process; The reagent storage module is used to store reagent containers containing reagents involved in the in vitro transcription process; The transfer module is used to receive the consumables and reagent containers transported by the robotic arm module; The robotic arm module is used to move the consumables from the consumables stack to the first target position of the transfer module based on the first handling command sent by the central controller, and to move the consumables at the first target position to the liquid workstation; The robotic arm module is used to move the reagent container from the reagent storage module to the second target position of the transfer module based on the second handling command sent by the central controller, and then move the reagent container at the second target position to the liquid workstation; The liquid workstation is used to receive pipetting instructions sent by the central controller and to perform pipetting operations on the consumables and reagent containers based on the pipetting instructions.

2. The in vitro transcription device according to claim 1, characterized in that, The transfer module is located between the consumables stack and the liquid workstation; And / or, the transfer module is made of aluminum alloy; And / or, the transfer module includes a top plate and a first side plate and a second side plate connected to and disposed opposite to the top plate, and a limiter is provided at the boundary of the top plate; And / or, the top plate of the transfer module is provided with a target area for accommodating the consumables or reagent containers of matching size.

3. The in vitro transcription device according to claim 1, characterized in that, The in vitro transcription device also includes a door opening device, the reagent storage module includes a refrigerator, and the reagents include low-temperature reagents; The refrigerator is used to store the low-temperature reagent container containing the low-temperature reagent; The door opening device is used to open the refrigerator door based on the door opening command sent by the central controller; The door opening device is also used to close the refrigerator door based on a door closing command sent by the central controller.

4. The in vitro transcription device according to claim 3, characterized in that, The door opening device includes a motor, a reduction mechanism, a connecting rod, a buckle, and a door opening controller; The buckle is connected to the door of the refrigerator, and the connecting rod, the reduction mechanism, and the motor are electrically connected in sequence. The door opening controller is used to drive the motor to operate based on the door opening command or the door closing command sent by the central controller; The motor is used to drive the reduction mechanism to perform a deceleration movement, thereby driving the connecting rod to move; The connecting rod is used to drive the latch to open or close the refrigerator door.

5. The in vitro transcription device according to claim 3, characterized in that, The in vitro transcription device also includes a sealing and tearing device, a shaking module, a centrifuge, and a PCR instrument; The central controller is used to control the operation of the sealing and tearing device, the shaking module, the centrifuge, and the PCR instrument, respectively. The consumables include an initial PCR plate and a first pipette tip box containing a first pipette tip. The low-temperature reagent includes a plasmid template and a PCR reaction solution; The liquid workstation is used to receive the initial PCR plate, the first pipette tip box, the plasmid template, and the PCR reaction solution container containing the PCR reaction solution transported by the robotic arm module, and to perform pipetting operations based on the first pipetting command sent by the central controller to obtain a PCR plate containing the amplification reaction solution. The robotic arm module is also used to transport the PCR plate containing the amplification reaction solution to the sealing and tearing device via the transfer module for sealing operation, so as to obtain a sealed amplification PCR plate. The robotic arm module is also used to transport the sealed amplification PCR plate to the oscillation module via the transfer module for oscillation to obtain an amplified and mixed PCR plate. The robotic arm module is also used to transport the amplified and mixed PCR plate to the centrifuge via the transfer module for centrifugation to obtain the amplified centrifuged PCR plate. The robotic arm module is also used to transport the amplified centrifuged PCR plate to the PCR instrument via the transfer module for PCR operation to obtain the plasmid amplification product plate.

6. The in vitro transcription device according to claim 5, characterized in that, The in vitro transcription device also includes a heating module, and the low-temperature reagent also includes a pre-sealed IVT premix; The robotic arm module is also used to transport the plasmid amplification product plate to the centrifuge via the transfer module for centrifugation to obtain a transcription centrifuged PCR plate. The robotic arm module is also used to transport the transcription centrifuged PCR plate to the sealing and tearing device via the transfer module for tearing operation to obtain the transcription tearing PCR plate; The robotic arm module is also used to transport the transcription membrane-tearing PCR plate to the liquid workstation via the transfer module; The robotic arm module is also used to transport the container containing the IVT premixed liquid to the centrifuge via the transfer module for centrifugation, so as to centrifuge the IVT premixed liquid into centrifuged IVT premixed liquid; the robotic arm module is also used to transport the container containing the centrifuged IVT premixed liquid to the liquid workstation via the transfer module. The robotic arm module is also used to transport the initial PCR plate in the consumable stack to the liquid workstation via the transfer module; The liquid workstation is also used to receive the initial PCR plate, the first pipette tip box, and the container containing the centrifuged IVT premixed solution transported by the robotic arm module, and to perform pipetting operations based on the second pipetting command sent by the central controller to obtain a PCR plate containing IVT reaction solution. The robotic arm module is also used to transport the PCR plate containing IVT reaction solution to the sealing and tearing device via the transfer module for sealing operation, so as to obtain a sealed PCR plate containing IVT reaction solution. The robotic arm module is also used to transfer the sealed PCR plate containing IVT reaction solution to the shaking module via the transfer module for shaking operation, so as to obtain a mixed PCR plate containing IVT reaction solution. The robotic arm module is also used to transfer the mixed PCR plate containing IVT reaction solution to the centrifuge via the transfer module for offline operation, so as to obtain a centrifuged IVT reaction solution PCR plate. The robotic arm module is also used to transport the centrifuged IVT reaction solution PCR plate to the heating module via the transfer module; The heating module is used to provide the corresponding first IVT reaction temperature to the centrifuged IVT reaction solution PCR plate based on the first IVT reaction temperature command sent by the central controller, so as to obtain the IVT product plate.

7. The in vitro transcription device according to claim 6, characterized in that, The low-temperature reagent includes a pre-sealed PCR plate containing DNase I; The robotic arm module is also used to transport the PCR plate containing DNase I to the sealing and tearing device via the transfer module for tearing operation, so as to obtain a PCR plate containing DNase I. The robotic arm module is also used to transport the DNase I-containing tear-off PCR plate to the liquid workstation via the transfer module; The robotic arm module is also used to transport the IVT product plate to the centrifuge via the transfer module for centrifugation to obtain centrifuged IVT product plate; The robotic arm module is also used to transport the centrifuged IVT product plate to the sealing and tearing machine via the transfer module for tearing operation to obtain the tearing IVT product plate. The robotic arm module is also used to transport the film-tear IVT product plate to the liquid workstation via the transfer module; The liquid workstation is used to receive the first pipette tip box, the membrane-tear IVT product plate and the membrane-tear PCR plate containing DNase I, and to perform pipetting operations based on the third pipetting command sent by the central controller to obtain a PCR plate containing DNase I digestion reaction solution. The robotic arm module is also used to transport the PCR plate containing DNase I digestion reaction solution to the sealing and tearing device via the transfer module for sealing operation, so as to obtain a sealed PCR plate containing DNase I digestion reaction solution. The robotic arm module is also used to transfer the sealed PCR plate containing DNaseI digestion reaction solution to the shaking module via the transfer module for shaking operation, so as to obtain a mixed PCR plate containing DNaseI digestion reaction solution. The robotic arm module is also used to transfer the mixed PCR plate containing DNase I digestion reaction solution to the centrifuge via the transfer module for centrifugation to obtain a centrifuged PCR plate containing DNase I digestion reaction solution. The robotic arm module is also used to transport the centrifuged PCR plate containing DNaseI digestion reaction solution to the heating module via the transfer module; The heating module is used to provide the corresponding second IVT reaction temperature to the centrifuged PCR plate containing DNaseI digestion solution based on the second IVT reaction temperature command sent by the central controller, so as to obtain the enzyme digestion product plate.

8. The in vitro transcription device according to claim 7, characterized in that, The robotic arm module is also used to transport the used DNase I-containing PCR plate to the sealing and tearing device via the transfer module for sealing operation. And / or, the robotic arm module is also used to transport the first pipette tip box containing the first pipette tip, located at the liquid workstation, to the consumables stack via the transfer module; And / or, the robotic arm module is also used to transport the enzyme digestion product plate to the refrigerator via the transfer module.

9. The in vitro transcription device according to claim 7, characterized in that, The reagent storage module includes a buffer rack, and the reagents include room temperature reagents; The buffer rack is used to store the room temperature reagent container containing the room temperature reagent; The robotic arm module is used to transport the room-temperature reagent container to the liquid workstation via the transfer module; Alternatively, the robotic arm module is used to transport the room-temperature reagent container to the buffer rack via the transfer module.

10. The in vitro transcription device according to claim 9, characterized in that, The in vitro transcription device also includes a magnetic bead purification device; The consumables include an initial deep-hole plate and a second pipette box equipped with a second pipette tip, wherein the pipetting capacity range of the second pipette tip is greater than that of the first pipette tip; The room-temperature reagent includes magnetic beads, binding buffer, washing buffer, and elution buffer; The robotic arm module is also used to transport the first liquid storage tank containing the magnetic beads, the second liquid storage tank containing the binding buffer, the third liquid storage tank containing the washing buffer, and the fourth liquid storage tank containing the elution buffer from the buffer rack to the liquid workstation via the transfer module, respectively. The robotic arm module is also used to transport the enzyme digestion product plate to the centrifuge via the transfer module for centrifugation to obtain the centrifuged enzyme digestion product plate. The robotic arm module is also used to transport the centrifuged enzyme digestion product plate to the sealing and tearing device via the transfer module for tearing operation to obtain the tearing enzyme digestion product plate. The robotic arm module is also used to transport the membrane-peeling enzyme digestion product plate and the initial deep well plate to the liquid workstation via the transfer module; The liquid workstation is also used to receive the initial deep well plate, the first pipette tip box, the second pipette tip box, the first liquid storage tank, the second liquid storage tank, the third liquid storage tank, the fourth liquid storage tank and the membrane-tear enzyme digestion product plate transported by the robotic arm module, and to perform pipetting operations based on the fourth pipetting command sent by the central controller to obtain a deep well plate for purification and enrichment of reagents. The robotic arm module is also used to transport the purification and enrichment reagent deep well plate to the magnetic bead purifier via the transfer module. The magnetic bead purifier is used to purify the purification enrichment reagent deep-well plate based on the purification instructions sent by the central controller, so as to obtain the mRNA deep-well plate.

11. The in vitro transcription device according to claim 10, characterized in that, The robotic arm module is also used to transport the cleaned magnetic bead deep-well plate after the purification operation to the magnetic bead purifier via the transfer module. The liquid workstation is also used to receive the first suction tip box, the second suction tip box, the first liquid storage tank, the second liquid storage tank, the third liquid storage tank, the fourth liquid storage tank and the cleaning magnetic bead deep well plate transported by the robotic arm module, and to perform a liquid transfer operation based on the fifth liquid transfer command sent by the central controller to obtain the magnetic bead recovery reagent deep well plate. The robotic arm module is also used to transport the magnetic bead recovery reagent deep well plate to the magnetic bead purifier via the transfer module; The magnetic bead purifier is used to perform a recovery operation on the cleaned magnetic beads in the magnetic bead recovery reagent deep well plate based on the recovery command sent by the central controller, so as to obtain a recovered magnetic bead deep well plate containing the recovered magnetic beads.

12. The in vitro transcription device according to claim 11, characterized in that, The robotic arm module is also used to transport the mRNA deep well plate or the recycled magnetic bead deep well plate to the buffer rack via the transfer module; And / or, the second pipette tip has a pipetting capacity range of 300 μL, and the first pipette tip has a pipetting capacity range of 20 μL.

Images