Transcription and translation coupled acellular teaching kit

Abstract

The utility model discloses a transcription and translation coupled acellular teaching kit. The transcription and translation coupled acellular teaching kit comprises a kit body, a reaction observation module, a heating module and a circuit module are arranged in the kit body, the heating module is connected with the reaction observation module, the circuit module is connected with the heating module, a reaction window is arranged on the side wall of the kit body, the reaction window is communicated with the reaction observation module, and the circuit module is connected with the reaction observation module. The reaction observation module is used for observing the reaction process in the reaction observation module. The reaction observation module ingeniously integrates reaction and observation functions, and through the integrated design, the whole using process becomes simple, rapid and easy to operate. And the circuit module is directly integrated in the kit body, so that the overall size is obviously reduced, and the portability is improved. Meanwhile, the integrated design also effectively reduces the production cost.

Description

Technical Field

[0001] This invention relates to the field of reagent kit technology, and more particularly to a cell-free teaching reagent kit for transcription-translation coupling. Background Technology

[0002] Current technologies have successfully enabled the expression and production of large quantities of proteins. However, due to various factors, some biologically significant proteins, such as cytotoxic proteins and poorly soluble proteins, remain difficult to express efficiently. With the continuous advancement of science and technology, cell-free protein expression technology, due to its unique open system characteristics, can precisely control reaction conditions, regulate reactant composition, and achieve direct monitoring of the reaction process, thus becoming a powerful protein expression method.

[0003] Cell-free protein synthesis (CFPS) systems, as an in vitro gene expression technology, utilize exogenous DNA or mRNA as templates and synthesize proteins in a cell-extracted environment by artificially supplementing essential raw materials and energy substances. This system can overcome the inherent limitations of cells, enabling rapid and convenient expression of a variety of proteins. Specifically, the CFPS system uses DNA as a template, and under the combined action of components such as RNA polymerase and transcription factors, a transcription process occurs to produce the corresponding mRNA. Subsequently, using mRNA as a template, and with the help of ribosomes, amino acid substrates, tRNA, and energy substances within the system, protein translation and synthesis are completed. Currently, core concepts in synthetic biology, such as protein expression, mainly exist in textbooks in theoretical form. Students often find these concepts abstract and vague, leading to a lack of interest and difficulty in understanding them, thus affecting their learning efficiency.

[0004] In the existing technology, there is a kit called BioBits™ Bright for synthetic biology education. However, this kit is expensive, which may reduce the number of repeated experiments, affecting data accuracy and reliability, and increasing the risk of experimental failure. Furthermore, the kit is designed for use by 30 students, which may reduce the quality of each student's individual experience during the experiment.

[0005] Therefore, there is an urgent need for a more convenient, simple, and lower-cost small-scale cell-free reaction teaching kit. Utility Model Content

[0006] The main objective of this invention is to provide a cell-free teaching kit for transcription-translation coupling, thereby overcoming the shortcomings of existing technologies.

[0007] To achieve the aforementioned objectives, the technical solution adopted by this utility model includes:

[0008] This invention provides a cell-free teaching kit for transcription-translation coupling, comprising a kit body, a reaction observation module, a heating module, and a circuit module. The reaction observation module, the heating module, and the circuit module are encapsulated inside the kit body. The heating module is connected to the reaction observation module, and the circuit module is connected to the heating module. A reaction window is provided on the side wall of the kit body, which communicates with the reaction observation module for observing the reaction process within the reaction observation module.

[0009] In some more specific implementations, the reaction observation module includes a reaction chamber, the reagent kit has multiple reaction holes that communicate with the reaction chamber, a reaction tube passes through the reaction holes and is placed inside the reaction chamber, and / or, the reaction tube passes through the reaction holes, with one end of the reaction tube fixed to the reaction hole and the other end placed inside the reaction chamber.

[0010] In some more specific implementations, the heating module includes a heating chamber, and a heat conduction structure is provided between the heating chamber and the reaction chamber, through which heat in the heating chamber is input into the reaction chamber.

[0011] Specifically, in reaction experiments, the heating element in the heating module often leads to high temperatures in localized areas, which may cause the reaction components to degrade. To address this issue, using a heating chamber is a gentler and more suitable method, especially considering that biological reactions often involve sensitive biomolecules. This heating chamber is connected to the reaction chamber via a heat conduction structure, ensuring that heat from the heating chamber can be effectively transferred to the reaction chamber, thus providing the necessary thermal energy for the reaction within the reaction chamber. More specifically, the heat conduction structure may include multiple heat conduction holes. These heat conduction holes are arranged to maximize heat transfer efficiency while maintaining thermal isolation between the heating chamber and the reaction chamber to avoid unnecessary heat loss. Furthermore, the multiple heat conduction holes are distributed in an array; this array layout helps to evenly distribute heat, ensuring a more uniform temperature within the reaction chamber.

[0012] In some more specific implementation schemes, the heating module further includes a heating element and a temperature control unit. The heating element is disposed inside the heating cavity. The heating element, the temperature control unit, and the circuit module are interconnected. The temperature control unit is used to monitor the temperature inside the heating cavity. When the temperature value reaches a preset temperature threshold, the temperature control unit will immediately perform a circuit breaker operation to prevent overheating.

[0013] The specific preset temperature threshold, temperature control unit, and heating element can be set according to the actual experiment.

[0014] Preferably, the preset temperature threshold can be set to 30℃. The temperature control unit can be implemented as a temperature switch, which can quickly respond and cut off the circuit when the set temperature is reached, thereby stopping the heating process. As for the heating element, an electric heating film can be selected, as it is widely used in various heating devices due to its high efficiency and fast response. Finally, the heating element is located on the bottom surface of the heating cavity; this arrangement helps to distribute heat evenly, thereby improving heating efficiency.

[0015] In some more specific implementations, a filter unit is provided on the reaction window to filter out the excitation light, ensuring that only light of a specific wavelength can pass through, thereby improving the accuracy and reliability of experimental data. Furthermore, these filter units can take various forms; for example, the filter unit can be a filter itself. This design is simple and effective, and can meet the needs of most experiments.

[0016] Preferably, the filter is an orange filter, which can reduce the interference of background blue excitation light while allowing the green fluorescence excited after successful expression to pass through, thereby improving the clarity and contrast of observation or imaging.

[0017] In some more specific implementations, the reaction observation module further includes an excitation light source. The excitation light source is located within the reaction chamber and connected to the circuit module, used to generate excitation light and act on the sample in the reaction tube. This excites specific substances in the sample to emit fluorescence, enabling further observation and analysis. Furthermore, the excitation light source is preferably positioned at the bottom of the reaction chamber to more uniformly illuminate the sample in the reaction tube.

[0018] Preferably, in the selection of light source type, blue LEDs can be selected, but are not limited to. Blue LEDs can excite a reaction to produce green fluorescent protein, thereby enabling clearer and more accurate observation.

[0019] In some more specific implementations, the circuit module includes a first circuit, a second circuit, and a power supply. The first circuit, the second circuit, and the power supply are connected in parallel. The first circuit is connected in series with the heating module, and the second circuit is connected in series with the excitation light source.

[0020] In some more specific implementations, the circuit module further includes a circuit cavity, a heating switch, a lighting switch, and a power switch. The heating switch is provided on the first circuit to allow the user to control the operating status of the heating module. Similarly, a lighting switch is provided on the second circuit to control the switching of the excitation light source. The power supply and the power switch are directly connected, allowing the user to turn the power to the entire circuit module on or off using the power switch. Furthermore, power supply holes, heating switch holes, and lighting switch holes are provided on the walls of the circuit cavity. These holes are designed for easy installation and fixation of the power switch, heating switch, and lighting switch, thereby ensuring the stability and safety of the entire circuit module.

[0021] In some more specific implementations, the reagent kit body also includes a circuit slot, which is used to accommodate and protect the first circuit and the second circuit. By placing the circuit slot inside the reagent kit body, external interference can be effectively isolated, while ensuring the stability and reliability of the circuit components.

[0022] Preferably, based on a comprehensive consideration of the performance and safety of the entire circuit module, the power supply is a 5V power supply. A 5V power supply can provide a stable voltage output, which not only meets the operating requirements of the circuit module, but also avoids the safety hazards that may be caused by excessively high voltage.

[0023] In some more specific implementations, the heating module and the circuit module are provided with a closed structure on top, and the closed structure is closely fitted to the heating module and the circuit module.

[0024] In some more specific implementations, the reagent kit includes a box body and a box cap, with the reaction reagent cap disposed on the top of the reagent kit body, the box cap being detachably disposed on the box body, and the reagent cap being disposed on the top of the closed structure.

[0025] Compared with the prior art, the advantages of this utility model include at least the following:

[0026] First, in the transcription-translation coupling cell-free teaching kit provided by this invention, the circuit module is directly integrated into the kit body, significantly reducing the overall size and increasing portability. At the same time, this integrated design also effectively reduces production costs.

[0027] Secondly, in the cell-free teaching kit for transcription-translation coupling provided by this utility model, a heat conduction structure is set between the heating chamber and the reaction chamber, so that the heat in the heating chamber can be effectively transferred to the reaction chamber through heat conduction. This ensures that the heat energy in the heating chamber can be transferred to the reaction chamber to the maximum extent through the heat conduction structure, thereby avoiding unnecessary heat loss during the transfer process and improving the thermal efficiency of the entire system.

[0028] Third, in the transcription-translation coupling cell-free teaching kit provided by this utility model, the heating element, temperature control unit and circuit module are connected. The temperature control unit continuously monitors the temperature change in the heating chamber. Once the temperature value is detected to reach the preset temperature threshold, the temperature control unit will react quickly and perform a circuit breaker operation to prevent the heating module from overheating due to excessive temperature, thereby ensuring the safe operation of the equipment.

[0029] Fourth, in the transcription-translation coupling cell-free teaching kit provided by this invention, the circuit cavity is a structural space for accommodating the circuit module, providing a closed environment to protect the internal circuit from external environmental influences. Heating switches, lighting switches, and power switches are respectively located on the first circuit, second circuit, and power supply. In this way, users can flexibly control the circuit from the outside, which not only improves the ease of use of the circuit module but also enhances operational safety. Attached Figure Description

[0030] Figure 1 This is a schematic diagram of the structure of a cell-free teaching kit for transcription-translation coupling provided in an embodiment of this utility model;

[0031] Figure 2 This is a schematic diagram of the structure of a cell-free teaching kit for transcription-translation coupling provided in an embodiment of this utility model;

[0032] Figure 3 This is a cross-sectional schematic diagram of a transcription-translation coupling cell-free teaching kit provided in an embodiment of this utility model;

[0033] Figure 4 This is a cross-sectional schematic diagram of a transcription-translation coupling cell-free teaching kit provided in an embodiment of this utility model.

[0034] Explanation of reference numerals in the attached figures:

[0035] 1. Reagent kit; 11. Reaction chamber; 12. Reaction well; 13. Filter; 14. Excitation source; 21. Heating chamber; 22. Heat conduction hole; 23. Heating element; 31. Circuit chamber; 32. Heating switch hole; 33. Heating switch; 34. Power supply hole; 41. Sealing sheet; 42. Reaction kit cap; 43. Cap groove; Detailed Implementation

[0036] In view of the shortcomings of the prior art, the inventor of this case, through long-term research and extensive practice, has come up with the technical solution of this utility model. The following will further explain the technical solution, its implementation process, and its principles.

[0037] Please refer to Figures 1-4A cell-free teaching kit for transcription-translation coupling includes a kit body 1, which internally comprises a reaction chamber 11, a heating chamber 21, and a circuit chamber 31. The reaction chamber 11 and the heating chamber 21 are tightly connected to ensure effective heat transfer, and the circuit chamber 31 is connected to the heating chamber 21. A reaction window is provided on the side wall of the kit body 1, which communicates with the reaction chamber 11, allowing the user to visually observe the reaction process within the reaction chamber 11 and thus monitor the experimental process in real time.

[0038] In this embodiment, the reagent kit 1 is provided with multiple reaction holes 12, which allow the reaction tube to pass through. One end of the tube is fixed to the reaction hole 12, and the other end is placed inside the reaction chamber 11. To improve the observation effect, a filter 13 is provided on the reaction window. This filter 13 can effectively filter out the excitation light and reduce interference with the observation. In addition, an excitation light source 14 is also included. This excitation light source 14 is located at the bottom of the reaction chamber 11 and connected to the circuit chamber 31. It is used to generate excitation light and act on the reaction tube, thereby exciting the sample inside the reaction tube to produce fluorescence, which is convenient for observation.

[0039] In this embodiment, multiple heat conduction holes 22 are provided between the heating chamber 21 and the reaction chamber 11. These heat conduction holes 22 can effectively transfer heat from the heating chamber 21 to the reaction chamber 11, ensuring that the reaction process takes place at a suitable temperature. This solution also includes a heating element 23 and a temperature control unit. The heating element 23 is disposed on the bottom surface of the heating chamber 21 and is interconnected with the temperature control unit. The temperature control unit is responsible for monitoring the temperature inside the heating chamber 21. When the temperature reaches a preset temperature threshold, the temperature control unit automatically disconnects the circuit and stops heating to prevent overheating. Here, the heating element 23 is disposed on the bottom surface of the heating chamber 21 to ensure uniform heat distribution.

[0040] This embodiment includes a first circuit, a second circuit, a power supply, a heating switch 33, a lighting switch, and a power switch. The first circuit and the second circuit are connected in parallel with the power supply. The heating switch 33 is installed on the first circuit, and the lighting switch is installed on the second circuit, ensuring independent control of heating and excitation light. The power supply is connected to the power switch. The wall of the circuit cavity 31 is provided with a power supply hole 34, a heating switch hole 32, and a lighting switch hole, which are used to install the power switch, heating switch 33, and lighting switch, respectively. In this way, various operations can be easily performed from the outside of the reagent kit 1, making the whole process simpler and more intuitive.

[0041] To further optimize the internal structure of the reagent kit 1, a circuit slot is provided inside the kit 1 to accommodate the first circuit and the second circuit, ensuring the stability and safety of the circuit. Furthermore, to protect the internal components and prevent interference from external factors, a sealing plate 41 is provided on the top of the heating chamber 21 and the circuit chamber 31. This sealing plate 41 fits tightly against the heating chamber 21 and the circuit chamber 31, ensuring the kit's airtightness and the stability of the internal environment.

[0042] Finally, a reaction kit cap 42 is provided on the top of the kit body 1. The reaction kit cap 42 is fixed to the kit body 1 by two corresponding cap slots 43, which can reduce the leakage of blue light excitation. In addition, after all experimental steps are completed, simply close the reaction kit cap 42 carefully. This not only effectively prevents dust and other contaminants from entering the kit, maintaining the purity of the reagents and the accuracy of experimental results, but also greatly improves the portability and storage convenience of the kit. The capped kit can be easily placed anywhere in or outside the laboratory, making it convenient for users to organize and store the kit after the experiment, ensuring the cleanliness and orderliness of experimental equipment.

[0043] Example

[0044] The method of using a cell-free teaching kit for transcription-translation coupling includes the following steps.

[0045] 1. Preparation stage:

[0046] Before using the transcription-translation coupling cell-free teaching kit, you must first prepare all the components and reagents required for the cell-free protein expression reaction according to the detailed requirements in the instruction manual.

[0047] This kit contains three main reagents, labeled A, B, and C. Reagent A consists of 57 mM MEPES-KOH buffer, 1.5 mM ATP, 1.5 mM UTP, 1.5 mM CTP, 1.5 mM GTP, 0.64 mM cAMP, 100 mM potassium glutamate, 12 mM ammonium acetate, 15 mM magnesium acetate, 2 mM amino acids, 2% (w / w) PEG8000, 33 mM phosphoenolpyruvate, and 25% (v / v) E. coli extract. Reagent B is purified deionized water; and Reagent C is a GFP plasmid containing a specific template sequence.

[0048] 2. Reaction stage:

[0049] After the preparation stage is completed, the reaction stage begins. First, reagents A and B, or reagents A and C, need to be mixed according to the experimental design. The mixed reagents are added to 1.5 mL centrifuge tubes to form two experimental groups: a negative control group without template and a positive control group with template. These two centrifuge tubes are placed on reaction well 12. Then, heating switch 33 is turned on to start the heating program, causing reaction chamber 11 to heat up. After approximately 2 hours, once the reaction is complete, heating switch 33 is turned off, and the illumination switch is immediately turned on to observe the reaction. Observation clearly shows that no obvious fluorescence signal appears in the negative control group, while a clear fluorescence signal is visible in the positive control group. This indicates that the template successfully expressed green fluorescent protein.

[0050] It should be understood that the above embodiments are merely illustrative of the technical concept and features of this utility model, and are intended to enable those skilled in the art to understand the content of this utility model and implement it accordingly. They should not be construed as limiting the scope of protection of this utility model. All equivalent changes or modifications made in accordance with the spirit and essence of this utility model should be included within the scope of protection of this utility model.

Claims

1. A cell-free educational kit for transcription translation coupling, characterized in that, The kit body, the reaction observation module, the heating module and the circuit module are packaged inside the kit body, the heating module is connected with the reaction observation module, the circuit module is connected with the heating module, a reaction window is arranged on the side wall of the kit body, the reaction window is communicated with the reaction observation module, and the reaction process in the reaction observation module is observed. The reaction observation module comprises a reaction cavity, a plurality of reaction holes are arranged on the kit body, the reaction holes are communicated with the reaction cavity, a reaction tube passes through the reaction hole and is arranged in the reaction cavity, and / or the reaction tube passes through the reaction hole, one end of the reaction tube is fixed to the reaction hole, and the other end is arranged in the reaction cavity.

2. The transcription-translation coupled cell-free educational kit of claim 1, wherein, The heating module comprises a heating cavity, a heat conduction structure is arranged between the heating cavity and the reaction cavity, heat in the heating cavity is input into the reaction cavity through the heat conduction structure, 3. The transcription-translation coupled cell-free educational kit of claim 2, wherein, and / or the heat conduction structure comprises a plurality of heat conduction holes; and / or the heat conduction holes are arranged in an array. The heating module further comprises a heating body and a temperature control unit, the heating body is arranged in the heating cavity, the heating body, the temperature control unit and the circuit module are connected with each other, the temperature control unit is used for monitoring the temperature in the heating cavity, and when the temperature value reaches a preset temperature threshold, the temperature control unit is disconnected, 4. The transcription-translation coupled cell-free educational kit of claim 3, wherein, and / or the heating body is arranged on the bottom surface of the heating cavity. An optical filter unit is arranged on the reaction window, and the optical filter unit is used for filtering excitation light.

5. The transcription-translation coupled cell-free educational kit of claim 1, wherein, The reaction observation module further comprises an excitation light source, the excitation light source is arranged in the reaction cavity and connected with the circuit module, and is used for generating excitation light and acting on the reaction tube, 6. The transcription-translation coupled cell-free educational kit of claim 2, wherein, and / or the excitation light source is arranged at the bottom of the reaction cavity. The circuit module comprises a first circuit, a second circuit and a power supply, the first circuit and the second circuit are connected in parallel with the power supply, the first circuit is connected in series with the heating module, and the second circuit is connected in series with the excitation light source.

7. The transcription-translation coupled cell-free educational kit of claim 6, wherein, The circuit module further comprises a circuit cavity, a heating switch, an illumination switch and a power supply switch, the heating switch is arranged on the first circuit, the illumination switch is arranged on the second circuit, the power supply is connected with the power supply switch, and a power supply hole, a heating switch hole and an illumination switch hole are arranged on the wall surface of the circuit cavity and are used for mounting the power supply switch, the heating switch and the illumination switch respectively.

8. The transcription-translation coupled cell-free educational kit of claim 7, wherein, The kit body further comprises a circuit slot arranged inside, and the circuit slot is used for accommodating the first circuit and the second circuit.

9. The transcription-translation coupled cell-free educational kit of claim 7, wherein, The heating module and the circuit module are provided with a sealing structure on the top, the sealing structure is closely combined with the heating module and the circuit module, 10. The transcription-translation coupled cell-free educational kit of claim 1, wherein, and / or the kit body comprises a box body and a box cover, and the box cover is arranged on the box body in an openable mode. ​

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