A device for detecting rice gene expression and function verification

Abstract

The application discloses a rice gene expression and function verification integrated detection device, which comprises a box body, a liquid nitrogen freezing assembly for freezing rice is installed in the box body, a gene extraction assembly for extracting rice genes is installed in the box body, and a gene detection assembly for detecting and verifying rice genes is installed at the top of the box body; a plurality of steps such as liquid nitrogen freezing, sample grinding, nucleic acid extraction, purification and qPCR detection are integrated, full-process automatic operation is realized through a control module, manual frequent intervention is not needed, human operation errors are greatly reduced, the accuracy and repeatability of detection results are improved, a high-concentration saline solution is combined, a low-salt elution process design is adopted, automatic magnetic bead adsorption and cleaning processes are combined, interference substances such as proteins and impurities in samples are effectively removed, the nucleic acid purity can reach more than 95%, the recovery rate can reach more than 85%, and the reliability of gene expression and function verification results is ensured.

Description

Technical Field

[0001] This invention relates to the field of rice gene detection technology, specifically to an integrated detection device for rice gene expression and function verification. Background Technology

[0002] As one of the most important food crops, rice's yield and quality directly affect food security and sustainable agricultural development. With the rapid development of molecular biology and genomics, precise verification of rice gene expression and function has become a core aspect of rice genetic breeding, variety improvement, and stress resistance research. Nucleic acid extraction and purification are fundamental to gene detection, and their quality and efficiency directly determine the reliability of subsequent gene expression analysis and functional verification results.

[0003] Currently, the mainstream process for rice gene testing in the industry still relies primarily on a combination of traditional manual operation and separate equipment. In this model, steps such as freezing, grinding, nucleic acid extraction, purification, and subsequent qPCR testing of rice samples must be completed step-by-step in separate devices with different functions, and a large amount of operation depends on manual intervention. Specifically, the traditional testing process has the following technical shortcomings: First, the process is cumbersome and involves high costs for manual intervention. Starting with sample pretreatment, the sample must first be frozen in liquid nitrogen, then transferred to a grinder to pulverize the tissue, followed by manual pipetting to centrifuge tubes for lysis, nucleic acid adsorption, and washing. Finally, the purified nucleic acid sample is transferred to a qPCR instrument for concentration and purity testing. The entire process involves multiple equipment switches and manual operations, which is not only time-consuming but also prone to problems such as manual pipetting errors and sample cross-contamination, resulting in poor repeatability and insufficient accuracy of the test results.

[0004] Secondly, the equipment has low integration and occupies a large space. Existing testing solutions require multiple independent instruments, such as liquid nitrogen freezing equipment, grinders, nucleic acid extraction devices, and qPCR instruments. The equipment is scattered and has poor compatibility, which not only increases the space occupied in the laboratory and the cost of equipment procurement, but also makes it impossible to achieve intelligent management and control of the testing process due to the lack of data communication between the devices. This makes it difficult to meet the rapid testing needs of large batches of rice samples in modern agriculture.

[0005] Therefore, developing a detection device that integrates rice sample freezing and grinding, automatic nucleic acid extraction and purification, gene expression and functional verification to achieve fully automated, high-precision and high-efficiency detection has become an urgent need to solve the pain points of existing technologies and promote the development of rice molecular breeding technology. Summary of the Invention

[0006] The purpose of this invention is to provide an integrated detection device for rice gene expression and function verification, so as to solve the problems mentioned in the background art.

[0007] To achieve the above objectives, the present invention provides the following technical solution: an integrated detection device for rice gene expression and function verification, comprising a box, wherein a liquid nitrogen freezing component for freezing rice is installed inside the box, a gene extraction component for extracting rice genes is installed inside the box, and a gene detection component for detecting and verifying rice genes is installed on the top of the box.

[0008] Preferably, the liquid nitrogen freezing assembly includes a liquid nitrogen freezing chamber, the interior of which has a freezing placement slot, a freezing chamber handle is fixedly installed on one side of the liquid nitrogen freezing chamber, and the liquid nitrogen freezing chamber is movably installed at the bottom of one side of the housing.

[0009] Preferably, the gene extraction component includes a placement component and a cleaning and drainage component. The placement component includes a placement chamber, into which a test tube is fixedly inserted, and on one side of the placement chamber is a handle for picking up and putting down.

[0010] Preferably, the placement compartment has grooves on both sides, the placement compartment is slidably installed inside the box, and a slider is fixedly installed inside the box, the slider is slidably installed inside the groove.

[0011] Preferably, the cleaning and drainage assembly includes a cleaning assembly and a liquid extraction assembly. The cleaning assembly includes a diversion box, the bottom of which is connected to multiple buffer cylinders, and the bottom of each of the multiple buffer cylinders is connected to a cleaning nozzle.

[0012] Preferably, the diversion box is fixedly installed inside the housing, and the top of the diversion box is connected to two inlet pipes, which are a high-concentration saline pipe and a low-concentration saline pipe, respectively.

[0013] Preferably, the liquid extraction assembly includes a movable plate, a supporting partition fixedly installed on the top of the movable plate, a first collecting pipe fixedly installed on the top of the supporting partition, a suction pipe connected to the bottom of each of the first collecting pipes, an electromagnet fixedly installed on the outer side of each suction pipe, a second collecting pipe connected to one end of each of the first collecting pipes, a third collecting pipe connected to one side of each of the second collecting pipes, a fourth collecting pipe connected to one side of each of the third collecting pipes, an outlet pipe connected to one side of the fourth collecting pipe, an external suction pipe connected to the end of the outlet pipe away from the fourth collecting pipe, and the end of the external suction pipe away from the outlet pipe located on the outer side of the housing.

[0014] Preferably, both ends of the movable plate are fixedly installed with sliding plates, and the movable plate and the sliding plates are slidably installed inside the housing. An electric cylinder is installed on the top of one of the sliding plates, and the electric cylinder is fixedly installed inside the housing. A limiting slide post is fixedly installed inside the housing. The other sliding plate has a sliding hole inside, and the sliding plate is slidably installed on the outside of the limiting slide post through the sliding hole.

[0015] Preferably, the gene detection component includes a qPCR instrument, an extraction head is slidably mounted on the bottom of the qPCR instrument, and a placement frame is fixedly mounted on the bottom of the qPCR instrument.

[0016] Preferably, a control display is fixedly installed on one side of the qPCR instrument, a printer is fixedly installed on the other side of the qPCR instrument, and a printing display screen is fixedly installed on the top of the printer.

[0017] Compared with the prior art, the beneficial effects of the present invention are: through a highly integrated structural design, multiple originally separate technical steps such as liquid nitrogen cryogenic freezing of rice leaf samples, tissue grinding and crushing, nucleic acid adsorption and binding, impurity cleaning and purification, and qPCR fluorescence quantitative detection are integrated into a continuous closed-loop detection system. From sample pretreatment to final nucleic acid analysis, it can be completed in one stop, without the need to repeatedly transfer samples between multiple devices, frequently change consumables, and manually operate. The entire process is driven by a unified control module, which automatically completes actions such as timing control, reagent dispensing, magnetic separation, liquid aspiration, and temperature control. This significantly reduces experimental errors caused by manual pipetting, manual sample addition, and repeated cap opening, and minimizes sample cross-contamination, operational errors, and environmental interference. As a result, it greatly improves the stability, accuracy, and parallel repeatability of the test results, providing a reliable data foundation for rice gene expression quantification and functional verification. In addition, a magnetic bead separation strategy is adopted, which uses specific binding in a high-salt environment and gentle elution in a low-salt environment. Combined with a highly automated magnetic bead adsorption and multi-stage washing process, DNA / RNA released from rice samples can be efficiently and specifically adsorbed onto the surface of silica magnetic beads in a high-concentration saline system, while impurities such as proteins, polysaccharides, pigments and cell debris are effectively repelled and separated. Subsequently, non-specific adsorbed impurities are gradually removed by repeated rinsing with low-salt buffer, and nucleic acids are fully eluted from the surface of magnetic beads under low-salt conditions. In addition, the device's built-in precise temperature control, controllable flow rate spray, and programmable magnetic separation mechanism enable fine-tuning of binding time, washing intensity, and elution efficiency. This thoroughly removes interfering components such as protein impurities, phenolic substances, salt ions, and inhibitors from the sample. Practical verification has shown that this purification method can stably achieve a nucleic acid purity of over 95% and a nucleic acid recovery rate of no less than 85%. It can provide high-purity, high-integrity, and high-concentration high-quality nucleic acid templates for subsequent qPCR amplification and quantitative fluorescence detection, ensuring amplification efficiency and detection sensitivity from the source, and significantly improving the authenticity and reliability of rice gene expression level detection and gene function verification results. Attached Figure Description

[0018] Figure 1 This is a three-dimensional structural diagram of the present invention.

[0019] Figure 2 This is a three-dimensional structural schematic diagram from another perspective of the present invention.

[0020] Figure 3 This is a three-dimensional structural schematic diagram of the present invention from another perspective.

[0021] Figure 4 This is a partial three-dimensional structural diagram of the present invention.

[0022] Figure 5 This is a partial three-dimensional structural diagram of the present invention from another perspective.

[0023] Figure 6 This is a three-dimensional structural diagram of the liquid extraction component of the present invention.

[0024] Figure 7 This is a three-dimensional schematic diagram of the rinsing assembly of the present invention.

[0025] In the diagram: 1. Chamber; 2. Control display; 3. qPCR instrument; 4. Extraction head; 5. Placement frame; 6. Printer; 7. Printer display screen; 8. Liquid nitrogen freezing chamber; 9. Freezing chamber handle; 10. Pick-up and drop handle; 11. Placement chamber; 12. Test tube; 13. Slide groove; 14. Slider; 15. Sliding plate; 16. Electric cylinder; 17. Limiting slide column; 18. Moving plate; 19. Support partition; 20. Merging tube one; 21. Suction tube; 22. Merging tube two; 23. Merging tube three; 24. Merging tube four; 25. Discharge tube; 26. Electromagnet; 27. External suction tube; 28. Buffer cylinder; 29. ​​Cleaning nozzle; 30. Diversion box. Detailed Implementation

[0026] 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 embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0027] Please see Figures 1-6This invention provides a technical solution: an integrated detection device for rice gene expression and function verification, comprising a box 1, inside which is installed a liquid nitrogen freezing component for freezing rice, a gene extraction component for extracting rice genes, and a gene detection component for detecting and verifying rice genes. The liquid nitrogen freezing component includes a liquid nitrogen freezing chamber 8, inside which is a freezing placement groove. A freezing chamber handle 9 is fixedly installed on one side of the liquid nitrogen freezing chamber 8, and the liquid nitrogen freezing chamber 8 is movably installed at the bottom of one side of the box 1. The gene extraction component includes a placement component and a cleaning and drainage component. The placement component includes a placement chamber 11, inside which a test tube 12 is fixedly inserted. A pick-and-place handle 10 is fixedly installed on one side of the placement chamber 11, and sliding grooves 13 are provided on both sides of the placement chamber 11. The placement chamber 11 is slidably installed inside the box 1, and a slider 14 is fixedly installed inside the box 1, slidingly installed inside the sliding grooves 13.

[0028] The working principle of the above technical solution is as follows: Workers clean the collected fresh rice leaf samples to remove surface impurities and moisture. Then, the rice leaf samples are neatly placed on the sample rack inside the liquid nitrogen freezing chamber 8. The chamber door is closed, and the freezing time is preset to 30 minutes and the freezing temperature to -196℃ via the control panel on the outside of the device. After the preset freezing time is reached, the workers open the chamber door and remove the frozen rice leaf samples from the chamber. These samples are then transferred to an external grinding device for thorough grinding until the samples form a uniform powder, facilitating sufficient contact between the subsequent lysis buffer and the sample for fine grinding. Cell membrane rupture, protein removal, and DNA / RNA release: Lysis buffer (TE buffer containing 1% SDS) is added to the ground rice leaf sample powder at a ratio of 1:10 (g / mL) to ensure complete cell membrane rupture, protein denaturation and removal, and release of rice nucleic acid, thus completing the sample pretreatment step. The pretreated rice sample mixture is then slowly injected into the test tube 12 placed inside the placement chamber 11, ensuring that the sample mixture is completely submerged at the bottom of the test tube 12 and that the amount of sample added does not exceed 1 / 3 of the capacity of the test tube 12 to avoid liquid overflow caused by subsequent reagent addition.

[0029] In another implementation scheme, such as Figures 1-6As shown, the cleaning and drainage assembly includes a cleaning component and a suction component. The cleaning component includes a diversion box 30, with multiple buffer cylinders 28 connected to the bottom of the diversion box 30. Each buffer cylinder 28 has a cleaning spray pipe 29 connected to its bottom. The diversion box 30 is fixedly installed inside the housing 1. The top of the diversion box 30 is connected to two inlet pipes, one for high-concentration saline and the other for low-concentration saline. The diversion box 30 is a rectangular diversion cavity, fixedly installed above the placement chamber 11. The top of the diversion box 30 is connected via a flange. It includes high-concentration saline tubing and low-concentration saline tubing, both made of food-grade PP material and equipped with electromagnetic flow control valves. This allows for precise control of the injection flow rate and velocity of the high-concentration and low-concentration saline solutions, preventing excessive flow from impacting the magnetic beads and nucleic acid complexes and affecting nucleic acid extraction. The inlet ends of both tubing extend to the outside of the device body for connecting reagent storage bottles, enabling automated replenishment of the high-concentration and low-concentration saline solutions. The extraction assembly includes a moving plate 18. A support partition 19 is fixedly installed on the top of the device. A first converging pipe 20 is fixedly installed on the top of the support partition 19. The bottom of each converging pipe 20 is connected to a suction pipe 21. An electromagnet 26 is fixedly installed on the outside of each suction pipe 21. One end of each converging pipe 20 is connected to a second converging pipe 22. One side of each second converging pipe 22 is connected to a third converging pipe 23. One side of each third converging pipe 23 is connected to a fourth converging pipe 24. One side of the fourth converging pipe 24 is connected to an outlet pipe 25. The end of the outlet pipe 25 away from the fourth converging pipe 24 is connected to... An external suction pipe 27 is connected, with one end of the external suction pipe 27 away from the liquid outlet pipe 25 located on the outside of the housing 1. Both ends of the movable plate 18 are fixedly installed with sliding plates 15. The movable plate 18 and the sliding plates 15 are slidably installed inside the housing 1. An electric cylinder 16 is installed on the top of one of the sliding plates 15. The electric cylinder 16 is fixedly installed inside the housing 1. A limiting slide post 17 is fixedly installed inside the housing 1. The other sliding plate 15 has a sliding hole inside, and the sliding plate 15 is slidably installed outside the limiting slide post 17 through the sliding hole.

[0030] The control module opens the electromagnetic flow control valve of the high-concentration saline tube at the top of the distribution box 30, injecting high-concentration saline into the distribution box 30. The high-concentration saline has a concentration of 5 mol / L. Inside the distribution box 30, the high-concentration saline is diverted by a buffer baffle and flows evenly into the buffer cylinder 28. Then, it is sprayed into the test tube 12 through the cleaning nozzle 29 at the bottom of the buffer cylinder 28. Under high-salt conditions, the phosphate groups in the rice nucleic acid specifically bind to the silanol groups on the surface of the silica magnetic beads, causing the nucleic acid to adhere tightly to the surface of the magnetic beads, completing the initial binding of nucleic acid and magnetic beads. During this process, the temperature sensor monitors the internal temperature of the placement chamber 11 in real time. The control module automatically starts the heating module based on the temperature data to maintain the temperature inside the chamber at 37°C, increasing the temperature. The binding efficiency of nucleic acid and magnetic beads is assessed after initial binding. The control module activates the electric cylinder 16, whose output extends downwards, pushing the sliding plate 15 downwards along the lead screw. This, in turn, causes the moving plate 18 and the supporting partition 19 to move downwards synchronously. The supporting partition 19 then moves the collecting tube 20, the suction tube 21, and the electromagnet 26 downwards until the suction tube 21 and the electromagnet 26 are fully inserted into the test tube 12, with the bottom of the suction tube 21 5mm from the bottom of the test tube to avoid contact with the bottom of the test tube, which could lead to magnetic bead accumulation or suction tube blockage. The control module then supplies power to the electromagnet 26, causing it to generate magnetism and strongly attract the magnetic beads inside the test tube 12. This ensures the magnetic beads are completely attracted to the surface of the electromagnet 26, preventing them from flowing with the liquid. After the magnetic beads have been adsorbed, the control module starts the external liquid pump. The pump steadily extracts the liquid inside the test tube 12 through the external suction tube 27, the outlet tube 25, the four collecting tubes 24, the three collecting tubes 23, the two collecting tubes 22, and the suction tube 21 until all the liquid inside the test tube 12 is extracted, completing the drainage step. After drainage, the control module stops supplying power to the electromagnet 26, and the electromagnet 26 loses its magnetism. The magnetic beads adsorbed on the surface of the tube naturally fall to the bottom of the inner side of the test tube 12 under the action of gravity, completing the recovery of the magnetic beads. The control module then controls the electromagnetic flow control valve of the low-concentration saline tube at the top of the diversion box 30 to open, injecting low-concentration saline into the diversion box 30. The concentration of the low-concentration saline is 0.1 mol / L. Low-concentration saline solution flows sequentially through buffer cylinder 28 and cleaning nozzle 29, and is sprayed into the interior of test tube 12. This repeatedly washes the rice nucleic acid adsorbed on the surface of magnetic beads inside test tube 12. Under low-salt conditions, the binding force between rice nucleic acid and the surface of magnetic beads weakens, and the nucleic acid detaches from the surface of magnetic beads and dissolves in low-concentration saline solution. After rinsing and drainage are completed, the control module activates the retraction of electric cylinder 16. The output end of electric cylinder 16 retracts upward, driving sliding plate 15, moving plate 18, support partition 19, suction tube 21, and electromagnet 26 to move upward, so that suction tube 21 and electromagnet 26 are completely removed from the interior of test tube 12. At this time, the supernatant in test tube 12 is the purified DNA / RNA, completing the rice nucleic acid recovery step.

[0031] In another implementation scheme, such as Figures 1-6 As shown, the gene detection component includes a qPCR instrument 3. An extraction head 4 is slidably mounted on the bottom of the qPCR instrument 3. The qPCR instrument 3 uses quantitative real-time PCR detection technology and has a built-in high-precision temperature control system, enabling rapid amplification of nucleic acid samples and detection of fluorescence signals. The extraction head 4 is located on the top of the qPCR instrument 3. The extraction head 4 is a multi-channel extraction head that can simultaneously extract and detect nucleic acid samples from multiple test tubes 12, significantly improving detection efficiency. The extraction head 4 is size-compatible with the test tubes 12, allowing for precise insertion into the tubes to detect the concentration and purity of nucleic acid samples, with a detection accuracy of up to 0. The qPCR instrument has a capacity of 0.1 ng / μL, which meets the detection requirements for rice gene expression and function verification. A placement frame 5 is fixedly installed at the bottom of the qPCR instrument 3, a control display 2 is fixedly installed on one side of the qPCR instrument 3, and a printer 6 is fixedly installed on the other side of the qPCR instrument 3. A printing display screen 7 is fixedly installed on the top of the printer 6. The printing display screen 7 is used to preview the layout and content of the test report. As needed, the staff can control the printer 6 to print the test report through the printing display screen 7. The report content includes the concentration and purity of the nucleic acid sample, the amplification curve, and the results of gene expression quantitative analysis, which is convenient for the staff to archive and conduct subsequent analysis.

[0032] Staff remove the nucleic acid-collected test tube 12 from the placement chamber 11, gently wipe the outer wall of the tube to remove any dirt, and then place the test tube 12 stably inside the placement frame 5. The placement frame 5 has a porous frame structure, facilitating the orderly placement and handling of multiple test tubes 12. The control module activates the qPCR instrument 3, controlling the extraction head 4 to insert into the test tube 12 to detect the concentration and purity of the pure nucleic acid inside the tube. The qPCR instrument 3 uses the principle of fluorescence signal detection to accurately determine the OD260 / OD280 ratio of the nucleic acid sample, thereby determining the purity of the nucleic acid. Simultaneously, the concentration of nucleic acid is measured, and the test results are transmitted to the control display 2 in real time. Staff can visually view the concentration value, purity ratio, and detection curve of nucleic acid on the display. If staff need to archive or submit the test report, they can operate the control display 2 to start the printer 6. The print display screen 7 will preview the layout and content of the test report in real time. After confirming that there are no errors, the printer 6 will be controlled by the print display screen 7 to print out the test report. The report records in detail the concentration, purity, detection time, and device operating parameters of the nucleic acid sample, thus completing the entire rice gene expression and function verification detection process.

[0033] Working principle: Workers clean the collected fresh rice leaf samples to remove surface impurities and moisture. The samples are then neatly placed on a sample rack inside the liquid nitrogen freezing chamber 8. The chamber door is closed, and the freezing time is preset to 30 minutes and the freezing temperature to -196℃ via the control panel on the outside of the device. After the preset freezing time is reached, the chamber door is opened, and the frozen rice leaf samples are removed and transferred to an external grinding device. The samples are thoroughly ground until they form a uniform powder, facilitating contact between the lysis buffer and the sample, breaking cell membranes, removing proteins, and releasing DNA / RNA. The lysis buffer is then added to the ground rice leaf sample powder. The lysis buffer used was TE buffer containing 1% SDS, added at a ratio of sample powder to lysis buffer of 1:10 (g / mL) to ensure complete cell membrane rupture, protein denaturation and removal, and release of rice nucleic acid, thus completing the sample pretreatment step. The pretreated rice sample mixture was then slowly injected into the test tube 12 placed inside the placement chamber 11, ensuring the sample mixture was completely submerged at the bottom of the test tube 12, and that the sample addition did not exceed 1 / 3 of the test tube 12's capacity to prevent liquid overflow during subsequent reagent addition. The control module opened the electromagnetic flow control valve of the high-concentration saline tube at the top of the split chamber 30, injecting high-concentration saline into the split chamber 30. The high-concentration saline had a concentration of 5 mol / L. Inside the diversion box 30, the water is diverted by a buffer baffle and flows evenly into the buffer cylinder 28. Then, through the cleaning nozzle 29 at the bottom of the buffer cylinder 28, it is sprayed into the test tube 12. Under high-salt conditions, the phosphate groups in the rice nucleic acid specifically bind to the silanol groups on the surface of the silica magnetic beads, causing the nucleic acid to adhere tightly to the surface of the magnetic beads, completing the initial binding of nucleic acid and magnetic beads. During this process, the temperature sensor monitors the internal temperature of the placement chamber 11 in real time. The control module automatically starts the heating module based on the temperature data to maintain the temperature inside the chamber at 37°C, improving the binding efficiency of nucleic acid and magnetic beads. After the initial binding of nucleic acid is completed, the control module starts the electric cylinder 16. The output end of the electric cylinder 16 extends downward, pushing the sliding plate 15 to move downward along the lead screw, thereby driving the moving plate. 18 moves downwards synchronously with the support partition 19. The support partition 19 drives the first collecting tube 20, the suction tube 21, and the electromagnet 26 downwards until the suction tube 21 and the electromagnet 26 are fully inserted into the test tube 12, with the bottom end of the suction tube 21 5mm away from the bottom of the test tube to avoid touching the bottom of the test tube, which could cause the magnetic beads to accumulate or the suction tube to become blocked. The control module supplies power to the electromagnet 26, which generates magnetism and strongly attracts the magnetic beads inside the test tube 12, ensuring that the magnetic beads are completely attracted to the surface of the electromagnet 26 to prevent the magnetic beads from being lost with the liquid. After the magnetic beads are attracted, the control module starts the external liquid pump. The liquid pump pumps through the external suction tube 27, the outlet tube 25, the fourth collecting tube 24, the third collecting tube 23, the second collecting tube 22, and the suction tube 21.The liquid inside the test tube 12 is steadily extracted until all the liquid inside the test tube 12 is extracted, completing the drainage step. After drainage, the control module stops supplying power to the electromagnet 26, and the electromagnet 26 loses its magnetism. The magnetic beads adsorbed on the surface of the tube naturally fall to the bottom of the inner side of the test tube 12 under the action of gravity, completing the recovery of the magnetic beads. The control module then controls the electromagnetic flow control valve of the low-concentration saline tube at the top of the diversion box 30 to open, injecting low-concentration saline into the diversion box 30. The concentration of the low-concentration saline is 0.1 mol / L. The low-concentration saline flows sequentially through the buffer cylinder 28 and the cleaning nozzle 29, and is sprayed into the interior of the test tube 12, repeatedly rinsing the rice nucleic acid adsorbed on the surface of the magnetic beads inside the test tube 12. Under the given environment, the binding force between rice nucleic acid and the surface of magnetic beads weakens, causing the nucleic acid to detach from the magnetic beads and dissolve in low-concentration saline. After rinsing and drainage, the control module activates the retraction of electric cylinder 16. The output end of electric cylinder 16 retracts upward, driving the sliding plate 15, moving plate 18, support partition 19, suction tube 21, and electromagnet 26 upward, completely disengaging the suction tube 21 and electromagnet 26 from the inside of the test tube 12. At this point, the supernatant in the test tube 12 is the purified DNA / RNA, completing the rice nucleic acid recovery step. The staff removes the nucleic acid-recovered test tube 12 from the inside of the placement chamber 11, gently wipes the stains on the outer wall of the test tube, and then places the test tube 12 stably inside the placement frame 5. The frame 5 is a multi-porous frame structure, facilitating the orderly placement and handling of multiple test tubes 12. The control module activates the qPCR instrument 3, controlling the extraction head 4 to insert into the test tube 12 to detect the concentration and purity of the pure nucleic acid within the tube. The qPCR instrument 3 accurately measures the OD260 / OD280 ratio of the nucleic acid sample using the fluorescence signal detection principle, thereby determining the purity of the nucleic acid and simultaneously measuring its concentration. The test results are transmitted to the control display 2 in real time, allowing staff to visually view the nucleic acid concentration value, purity ratio, and detection curve. If staff need to archive or submit the test report, they can operate the control display 2 to activate the printer 6, and the print screen 7 will preview the layout and content of the test report in real time to confirm its accuracy. Subsequently, the printer 6 is controlled via the print display screen 7 to print out the test report. The report details the concentration, purity, detection time, and device operating parameters of the nucleic acid sample, completing the entire rice gene expression and function verification detection process. The device integrates multiple steps such as liquid nitrogen freezing, sample grinding, nucleic acid extraction, purification, and qPCR detection into one unit. The control module achieves fully automated operation, eliminating the need for frequent manual intervention, significantly reducing human error, and improving the accuracy and repeatability of the test results. The device employs a high-concentration saline solution combined with low-salt elution, along with automated magnetic bead adsorption and cleaning processes, effectively removing interfering substances such as proteins and impurities from the sample. The nucleic acid purity can reach over 95%, and the recovery rate can reach over 85%.This provided high-quality nucleic acid samples for subsequent qPCR testing, ensuring the reliability of gene expression and function verification results.

[0034] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. An integrated detection device for rice gene expression and function verification, comprising a housing (1), characterized in that: The box (1) is equipped with a liquid nitrogen freezing assembly for freezing rice, a gene extraction assembly for extracting rice genes, and a gene detection assembly for detecting and verifying rice genes.

2. The integrated detection device for rice gene expression and function verification according to claim 1, characterized in that: The liquid nitrogen freezing assembly includes a liquid nitrogen freezing chamber (8), which has a freezing placement slot inside. A freezing chamber handle (9) is fixedly installed on one side of the liquid nitrogen freezing chamber (8), and the liquid nitrogen freezing chamber (8) is movably installed on the bottom of one side of the box body (1).

3. The integrated detection device for rice gene expression and function verification according to claim 2, characterized in that: The gene extraction component includes a placement component and a cleaning and drainage component. The placement component includes a placement chamber (11), a test tube (12) is fixedly inserted inside the placement chamber (11), and a pick-and-place handle (10) is fixedly installed on one side of the placement chamber (11).

4. The integrated detection device for rice gene expression and function verification according to claim 3, characterized in that: The placement compartment (11) has sliding grooves (13) on both sides. The placement compartment (11) is slidably installed inside the box body (1). A slider (14) is fixedly installed inside the box body (1). The slider (14) is slidably installed inside the sliding groove (13).

5. The integrated detection device for rice gene expression and function verification according to claim 4, characterized in that: The cleaning and drainage assembly includes a cleaning assembly and a liquid extraction assembly. The cleaning assembly includes a diversion box (30), and the bottom of the diversion box (30) is connected to a plurality of buffer cylinders (28). The bottom of each of the plurality of buffer cylinders (28) is connected to a cleaning nozzle (29).

6. The integrated detection device for rice gene expression and function verification according to claim 5, characterized in that: The diversion box (30) is fixedly installed inside the box (1). The top of the diversion box (30) is connected to two inlet pipes, which are a high-concentration saline pipe and a low-concentration saline pipe, respectively.

7. The integrated detection device for rice gene expression and function verification according to claim 6, characterized in that: The liquid extraction assembly includes a movable plate (18), a support partition (19) is fixedly installed on the top of the movable plate (18), a first converging pipe (20) is fixedly installed on the top of the support partition (19), a suction pipe (21) is connected to the bottom of the first converging pipe (20), an electromagnet (26) is fixedly installed on the outside of the suction pipe (21), a second converging pipe (22) is connected to one end of the first converging pipe (20), a third converging pipe (23) is connected to one side of the second converging pipe (22), a fourth converging pipe (24) is connected to one side of the third converging pipe (23), an outlet pipe (25) is connected to one side of the fourth converging pipe (24), an external suction pipe (27) is connected to one end of the outlet pipe (25) away from the fourth converging pipe (24), and the end of the external suction pipe (27) away from the outlet pipe (25) is located outside the housing (1).

8. The integrated detection device for rice gene expression and function verification according to claim 7, characterized in that: Both ends of the movable plate (18) are fixedly installed with sliding plates (15). The movable plate (18) and the sliding plate (15) are slidably installed inside the housing (1). One of the sliding plates (15) is equipped with an electric cylinder (16) on its top. The electric cylinder (16) is fixedly installed inside the housing (1). The housing (1) is fixedly installed with a limiting slide post (17). The other sliding plate (15) has a sliding hole inside, and the sliding plate (15) is slidably installed on the outside of the limiting slide post (17) through the sliding hole.

9. The integrated detection device for rice gene expression and function verification according to claim 8, characterized in that: The gene detection component includes a qPCR instrument (3), an extraction head (4) is slidably mounted on the bottom of the qPCR instrument (3), and a placement frame (5) is fixedly mounted on the bottom of the qPCR instrument (3).

10. The integrated detection device for rice gene expression and function verification according to claim 9, characterized in that: A control display (2) is fixedly installed on one side of the qPCR instrument (3), and a printer (6) is fixedly installed on the other side of the qPCR instrument (3). A printing display screen (7) is fixedly installed on the top of the printer (6).

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