Simple and rapid multi-concentration glue preparation kit and glue preparation method

Abstract

The invention relates to the technical field of biochemical experiments, and particularly discloses a simple and rapid multi-concentration glue preparation kit and a glue preparation method.The simple and rapid multi-concentration glue preparation kit comprises lower-layer glue, upper-layer glue and a coagulant, and the upper-layer glue comprises an upper-layer glue solution A and an upper-layer glue solution B; the lower-layer glue A liquid comprises a lower-layer glue A1 liquid, a lower-layer glue A2 liquid and a lower-layer glue B liquid, the lower-layer glue A1 liquid, the lower-layer glue A2 liquid and ultrapure water are mixed to form the lower-layer glue A liquid with different concentrations, a user only needs to refer to a simple matching table, the lower-layer glue A1 liquid, the lower-layer glue A liquid and the ultrapure water are mixed according to different volumes, and the lower-layer glue A liquid and the lower-layer glue B liquid can be prepared on the basis of a general formula. The lower-layer separation gel working solution with various concentrations can be rapidly and accurately prepared, and the lower-layer gel solution B, the upper-layer gel solution A and the upper-layer gel solution B are kept fixed and universal, so that variable independent adjustment and constant unified supply are skillfully realized.

Description

Technical Field

[0001] This application relates to the field of biochemical experimental technology, and specifically discloses a simple and rapid multi-concentration gel preparation kit and gel preparation method. Background Technology

[0002] Polyacrylamide gel electrophoresis (SDS-PAGE) is a fundamental and crucial technique in biochemistry and molecular biology for the separation and identification of proteins. This technique is based on the principle that the charge of a protein in the presence of SDS (sodium dodecyl sulfate) is proportional to its molecular weight, and separation is achieved by migration within the polyacrylamide gel matrix according to molecular weight. The gel concentration (i.e., the total content of acrylamide monomers) is the core parameter determining its separation range: low-concentration gels (e.g., 8%) have larger pore sizes, which are beneficial for the separation of high molecular weight proteins; while high-concentration gels (e.g., 14%) have smaller pore sizes, which are more suitable for the effective resolution of low molecular weight proteins. Therefore, in practical research, it is often necessary to flexibly select different gel concentrations according to the molecular weight range of the target protein.

[0003] Currently, the preparation of SDS-PAGE gels in the laboratory mainly relies on the following methods: One method is the traditional manual preparation method: this requires researchers to independently weigh multiple reagents such as acrylamide, methylenebisacrylamide, Tris-HCl, and SDS, and precisely adjust the pH value. This method is cumbersome, takes 30-50 minutes, and requires a high level of skill from the operator. Inaccurate weighing or pH adjustment deviations can easily lead to large batch-to-batch variations and poor experimental repeatability. Furthermore, acrylamide monomers are neurotoxic, and frequent weighing increases the health risks for researchers.

[0004] Secondly, there are commercially available one-step reagent kits: companies like Yaxin offer products that simplify gel preparation. These products typically pre-mix or dispense multiple components from acrylamide / methylenebisacrylamide mixtures, buffers, initiators (such as ammonium persulfate), and accelerators (such as TEMED), reducing the workload for researchers in weighing and preparing multiple independent solutions, thus improving convenience and speed to some extent. However, these products usually only prepare one fixed concentration of gel per kit (e.g., only a 12% separating gel). When an experimental study requires the simultaneous analysis of multiple proteins with significantly different molecular weights, or when different experimental projects require gels with different separation ranges, researchers have to purchase and store multiple independent kits corresponding to different concentrations. This not only significantly increases reagent procurement costs but also occupies laboratory refrigerator or freezer storage space due to the need to store multiple packaging sizes of kits, resulting in a waste of resources and space.

[0005] As can be seen, current gel preparation kits, while allowing for rapid and flexible preparation of separating gels at different concentrations, lead to increased experimental time, reagent costs, and wasted reagent storage space. Therefore, this invention provides a simple and rapid multi-concentration gel preparation kit and method to address these problems. Summary of the Invention

[0006] The purpose of this invention is to solve the problem that traditional gel preparation kits, which quickly and flexibly prepare separating gels of different concentrations, lead to increased experimental operation time, reagent costs, and wasted reagent storage space.

[0007] To achieve the above objectives, the basic solution of the present invention provides a simple and rapid multi-concentration gel preparation kit, comprising a lower gel, an upper gel, and a coagulant, wherein the upper gel comprises upper gel A solution and upper gel B solution; The lower layer adhesive A solution includes lower layer adhesive A1 solution, lower layer adhesive A2 solution and lower layer adhesive B solution. The lower layer adhesive A1 solution and the lower layer adhesive A2 solution are mixed with ultrapure water to form lower layer adhesive A solutions of different concentrations.

[0008] Furthermore, the lower layer adhesive A1 solution is a mixed mother liquor of Acr and Bis; The lower layer gel A2 solution includes TEMED and ultrapure water; The lower layer adhesive B solution comprises Tris-HCl and SDS; The upper layer adhesive A solution includes Acr-Bis, TEMED, and ultrapure water; The upper layer of adhesive B solution includes Tris-HCl, SDS and ultrapure water; The coagulant is ammonium persulfate.

[0009] Furthermore, in the lower layer adhesive A1 solution, the concentration of the mixed mother liquor is 30%, and the ratio of Acr to Bis is 29:1; In the lower layer of adhesive A2 solution, the ratio of TEMED to ultrapure water is 0.16:12; In the lower layer of gel B solution, the pH of Tris-HCl is 8.8 and the SDS content is 10%. In the upper layer of adhesive A solution, the concentration of Acr-Bis is 30%, and the ratio of Acr-Bis, TEMED and ultrapure water is 34:0.2:66. In the upper gel B solution, the pH of Tris-HCl is 6.8, and the ratio of Tris-HCl, SDS and ultrapure water is 26:2:72. The concentration of ammonium persulfate in the coagulant is 10%.

[0010] Furthermore, the lower layer adhesive A solution is composed of lower layer adhesive A1 solution, lower layer adhesive A2 solution and ultrapure water mixed in the following volume ratio: 26:3.04:21.

[0011] Furthermore, the lower layer adhesive A solution is composed of lower layer adhesive A1 solution, lower layer adhesive A2 solution and ultrapure water mixed in the following volume ratio: 34:3.04:13.

[0012] Furthermore, the lower layer adhesive A solution is composed of lower layer adhesive A1 solution, lower layer adhesive A2 solution and ultrapure water mixed in the following volume ratio: 40:3.04:7.

[0013] Furthermore, the lower layer adhesive A solution is composed of lower layer adhesive A1 solution, lower layer adhesive A2 solution and ultrapure water mixed in the following volume ratio: 46:3.04:1.

[0014] Based on the same inventive concept, the present invention also provides a simple and rapid method for preparing separating gels of different concentrations, including using the above-mentioned gel preparation kit to prepare separating gels of different concentrations.

[0015] Furthermore, the steps for preparing separating gels of different concentrations are as follows: Step S1: Prepare the coagulant accelerator; Step S2: Sequentially measure the lower layer gel A1 solution, lower layer gel A2 solution, and ultrapure water to prepare the lower layer gel A solution of the corresponding concentration. Step S3: Take the lower layer adhesive A liquid, the lower layer adhesive B liquid and the coagulant and mix them to obtain the lower layer adhesive; take the upper layer adhesive A liquid and the upper layer adhesive B liquid and mix them with the coagulant to obtain the upper layer liquid. Step S4, Step S4, quickly pour the mixture into the gel mold and cover it with a layer of isopropanol to seal it; Step S5: Let stand at room temperature for 8-12 minutes. Observing a clear interface indicates that polymerization is complete.

[0016] Furthermore, the concentration of the lower layer adhesive A solution is 8%, 10%, 12%, or 14%.

[0017] The principle and effect of this solution are as follows: 1. Compared with existing technologies, this invention, through the design of lower layer gel A1 and lower layer gel A2 solutions, allows users to quickly and accurately prepare working solutions of various concentrations (e.g., 8%, 10%, 12%, 14%) of lower layer separating gels by simply referring to a simple mixing table and mixing lower layer gel A1 and lower layer gel A2 solutions with ultrapure water in different volumes, based on a universal formula. Meanwhile, lower layer gel B solution and upper layer gel A and upper layer gel B solutions remain fixed and universal. This ingeniously achieves independent adjustment of variables while maintaining a constant and unified supply.

[0018] 2. This invention allows a single reagent kit to cover commonly used concentration ranges through modular combinations, fundamentally solving the problem of fixed concentrations and the need for multiple kits to be stockpiled in commercial reagent kits. This significantly saves costs and space, and represents a breakthrough in multi-concentration flexibility.

[0019] 3. This invention retains the advantages of a one-step, weigh-free process while adding only one simple liquid mixing step to achieve concentration selection. The entire process eliminates contact with toxic powders, reducing the total mixing time to 8-12 minutes. It achieves an optimal balance between convenience and flexibility, optimizing both operational ease and safety.

[0020] 4. All key reagents in this invention are standardized premixed solutions, requiring only precise measurement of liquid volume for operation, greatly eliminating human error and ensuring high batch-to-batch consistency. One multi-concentration kit can replace multiple single-concentration products, significantly reducing unit experimental costs and offering excellent repeatability and economy.

[0021] 5. This invention allows for highly standardized operating procedures, making it easy for operators of varying experience levels to master. The optimized formulation and stable forms of key components (such as lyophilized coagulant powder) ensure the long-term storage stability and performance reliability of the kit, exhibiting excellent versatility and stability. Simultaneously, it reduces exposure to the toxic monomer acrylamide, lowering health risks for laboratory personnel and enhancing safety. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments of this application, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0023] Figure 1 A schematic diagram of a simple and rapid multi-concentration gel preparation kit according to an embodiment of this application is shown; Figure 2 Example images of the results of electrophoresis and membrane transfer using the kit are shown; Figure 3 An example image of GAPDH expression detected by Western blot using the kit is shown; Figure 4 This image shows an example of GAPDH expression detected by Western blot after the kit has been stored for 2 months. Figure 5 The following are examples of comparison between the kit and the 10% Yakult experiment, where (a) is an example of the 10% Yakult experiment and (b) is an example of the simple and rapid multi-concentration gel preparation kit proposed in the embodiments of this application. Detailed Implementation

[0024] To further illustrate the technical means and effects of the present invention in achieving its intended purpose, the following detailed description of the specific implementation methods, structures, features, and effects of the present invention, in conjunction with the accompanying drawings and preferred embodiments, is provided below.

[0025] A simple and rapid multi-concentration gel preparation kit, for example... Figure 1 As shown: This includes lower layer adhesive A1, lower layer adhesive A2, lower layer adhesive B, upper layer adhesive A, upper layer adhesive B, and a coagulant, as detailed below: The lower layer A1 solution is made by mixing 150 ml of a 30% concentration of acrylamide-methylenebisacrylamide in a 29:1 ratio. This concentration can cover the commonly used coagulation concentration range of 8%-15% through simple volume adjustment.

[0026] The lower gel A2 solution contains 160 μL of TEMED and 12 mL of ultrapure water.

[0027] The lower gel B solution is a separating gel buffer containing 1.5M Tris-HCl (pH 8.8) and 10% SDS. It is designed with a 2x concentration to ensure accurate final concentration after mixing with lower gel A of varying concentrations, while also reducing preparation steps.

[0028] The upper gel A solution contains 34 ml of 30% Acr-Bis, 0.2 ml of TEMED, and 66 ml of ultrapure water.

[0029] The upper gel B solution contains 26 ml of 1.0 M Tris-HCl (pH=6.8), 2 ml of 10% SDS and 72 ml of ultrapure water.

[0030] The coagulant is 10% ammonium persulfate, provided in lyophilized powder form, which ensures the stability and activity of the reagent.

[0031] In this embodiment, the experimental reagents are sourced as follows: 30% Acr / Bis (acrylamide / methylene) and ammonium persulfate powder were purchased from Biosharp. 1.5 M Tris-HCl (pH 8.8), 1.0 M Tris-HCl (pH 6.8) and TEMED (tetramethylethylenediamine) were purchased from Beyotime Biotechnology Co., Ltd. Polyacrylamide gel dye and protein rainbow marker were purchased from Wuhan Saiwei Co., Ltd. SDS, Tris-base and Glycine were purchased from Sigma.

[0032] The reagents are prepared as follows: 10% SDS: Weigh 1.0g of SDS using an electronic balance, add an appropriate amount of ultrapure water to a centrifuge tube, transfer the SDS into the centrifuge tube, vortex to fully dissolve the SDS powder, and then add ultrapure water to make the total volume 10ml.

[0033] 10% Ammonium Persulfate: Weigh 1.0g of ammonium persulfate powder using an electronic balance, add an appropriate amount of ultrapure water to a centrifuge tube, transfer the powder to the centrifuge tube, vortex to fully dissolve the ammonium persulfate powder, then add ultrapure water to make the total volume 10ml, dispense 1ml into EP tubes, label, and store at -20℃.

[0034] By mixing lower layer adhesive A1 and lower layer adhesive A2 in different proportions, lower layer adhesive A solutions of different concentrations can be prepared. Lower layer adhesive B solutions, upper layer adhesive A solutions, and upper layer adhesive B solutions are all the same and therefore can be used interchangeably.

[0035] The lower layer A1 solution is 30% Acr / Bis. Taking 150ml as an example, it can be subsequently prepared into 50ml each of 8%, 10%, 12% and 15% lower layer A solutions. Lower layer A2 solution is diluted TEMED. Take 12ml of ultrapure water, add 160ul of TEMED, and vortex mix well to prepare the lower layer A2 solution.

[0036] According to the table below, add the corresponding volumes of lower layer A1 solution, lower layer A2 solution, and ultrapure water in sequence to prepare lower layer A solution with four concentrations of 8%, 10%, 12.5%, and 15%.

[0037] lower layer adhesive A concentration Volume of lower gel A1 (ml) Volume of lower layer gel A2 (ml) Ultrapure water (ml) Total volume (ml) 8% 26 3.04 21 50.04 10% 34 3.04 13 50.04 12% 40 3.04 7 50.04 14% 46 3.04 1 50.04 Lower layer gel B solution: Prepare 200 ml of lower layer gel B solution by adding 104 ml of 1.5 M Tris-HCl (pH 8.8), 4 ml of 10% SDS and 92 ml of ultrapure water in sequence, mix well and store at 4°C for later use.

[0038] Upper layer A solution: Prepare 100ml of upper layer A solution by taking 34ml of 30% Acr-Bis, 0.2ml of TEMED and 66ml of ultrapure water in sequence, vortex to mix well and store in a brown bottle at 4℃ for later use.

[0039] Upper layer gel B solution: To prepare 100ml of upper layer gel B solution, add 26ml of 1.0M Tris-HCl (pH=6.8), 2ml of 10% SDS and 72ml of ultrapure water in sequence. Vortex mix well, then add 400ul of polyacrylamide gel dye, mix well and store at 4℃ for later use.

[0040] Accelerator: 10% ammonium persulfate is used as the accelerator, replacing the modified accelerator in the Yake one-step gel preparation kit.

[0041] Depending on the concentration of the lower layer gel A, to prepare separating gels of 8%, 10%, 12%, and 14%, first select the corresponding lower layer gel A, add the lower layer gel B at a 1:1 ratio, and then add an appropriate volume of coagulant. For example, to prepare 10 ml of separating gel, take 5 ml each of lower layer gel A and B, add 60 μl of 10% ammonium persulfate, mix well, and then add to a glass plate to prepare the lower layer gel.

[0042] Since the concentration of the stacking gel is the same, the same upper gel solution A and upper gel solution B are used. When preparing a 2ml stacking gel system, take 1ml of upper gel solution A and 1ml of upper gel solution B, and add 20ul of 10% ammonium persulfate. After vortexing and mixing, the upper gel is prepared.

[0043] To evaluate the electrophoresis performance of the kit in this embodiment, a Western blot experiment was performed: Cellular proteins were extracted using the RIPA lysis method, quantified using the BCA method, and then loaded with loading buffer. After vortexing and heating in a metal bath at 95 °C for 8 min, separating gels of 8%, 10%, 12%, and 14% were prepared as described above, and the surface of the separating gel was leveled with anhydrous ethanol. After solidification, the anhydrous ethanol was discarded, and a stacking gel was prepared. The stacking gel was added to the glass plate tank, and the electrophoresis comb was quickly inserted. The gel was allowed to stand at room temperature for 30 min until the boundary between the stacking and separating gels was clearly observed after solidification. The glass plate was then placed in the electrophoresis tank, and electrophoresis buffer was added to completely submerge the stacking gel. The electrophoresis comb was removed, and protein markers and samples were added. The electrophoresis apparatus was connected, and the parameters were set (voltage 80 V, adjusted to 120 V after the loading buffer passed through the stacking gel). The loading buffer was placed approximately 2 cm from the bottom of the glass plate. Transfer buffer was prepared, along with a PVDF membrane, filter paper, and methanol.

[0044] Turn off the electrophoresis apparatus, remove the glass plate from the electrophoresis tank, cut off the top of the stacking gel, and transfer the separating gel to the transfer buffer. Soak the filter paper in the transfer buffer, remove air bubbles with a roller, and transfer the moistened filter paper to the drawer. Activate the PVDF membrane with methanol and cover it with the filter paper. Then, place the separating gel and filter paper in sequence, remove air bubbles with a roller, and close the lid. Set the parameters and transfer the membrane (voltage: constant 23 V; time: 40 min). Block with rapid blocking buffer and incubate overnight at 4°C for GAPDH. The next day, wash the PVDF membrane three times with TBST buffer, 5 min each time. Prepare goat anti-mouse secondary antibody, place the PVDF membrane in the secondary antibody, and incubate on a shaker at room temperature for 2 h. Wash with TBST, prepare ECL developing solution (1:1, protected from light), and vortex to mix. Add developing solution to the PVDF membrane and observe the results using a gel imaging system.

[0045] The experimental results are as follows: Electrophoresis and transfer results: After electrophoresis, the protein rainbow markers were observed to be separated, with varying spacing between different gels. As the separating gel concentration increased, the distance between small protein markers gradually increased. In the 8% separating gel, the 15kDa and 10kDa protein markers overlapped, while the 70-160kDa protein markers had a larger spacing. In the 10% separating gel, the 15kDa and 10kDa markers had separated. The distance increased in the 12% and 14% gels. A similar pattern was observed in the PVDF membrane. Therefore, an 8% separating gel is recommended for Western blot experiments on large proteins, while 12% or 14% separating gels should be considered for small target proteins. Exposure results showed that all four different separating gel concentrations could effectively detect GAPDH expression.

[0046] To further evaluate the stability of the kit, the kit was stored at 4°C for 2 months, and electrophoresis and transfer were performed again using 10% and 12% separating gels. The results showed that GAPDH expression could be detected using the above kit, indicating that the rapid gel preparation kit is stable.

[0047] To assess the reliability of this product, the 10% YARN-T1 one-step gel preparation kit and the kit described in this example were compared. The results show that... Figure 5 As shown, both kits can detect GAPDH expression, indicating that the rapid gel mixing kit proposed in this embodiment is no less effective than commercial products.

[0048] Based on the same inventive concept, another embodiment of the present invention provides a simple and rapid method for preparing separating gels of different concentrations, including using the gel preparation kit of the above embodiment to prepare separating gels of different concentrations, the steps of which are as follows: Step S1: Take one tube of coagulant ammonium persulfate dry powder, add 100 μL of the matching diluent, and vortex to dissolve to obtain a 10% ammonium persulfate solution; Step S2: According to the preparation instructions, measure the lower layer gel A1 solution, lower layer gel A2 solution and ultrapure water in sequence to prepare 50.04 ml of lower layer gel A solution of various concentrations. Step S3: Take 2.7 ml each of lower layer gel A and lower layer gel B of different concentrations, and 60 μL of 10% APS solution, and vortex to mix. Step S4: Quickly pour the mixture into the gel mold and cover it with a layer of isopropanol to seal it. Step S5: Let stand at room temperature for 8-12 minutes. Observing a clear interface indicates that polymerization is complete.

[0049] To verify the performance of this embodiment, a 10% concentration separating gel was used to perform a Western blot experiment on GAPDH.

[0050] Experimental group: 10% separating gel was prepared using the kit of this invention.

[0051] Control group: Yamei one-step method 10% gel preparation kit.

[0052] Operation time: The average glue preparation time for the experimental group was 10 minutes; the average preparation time for the control group was 10 minutes.

[0053] Gel performance: All gels were used for electrophoresis of standard protein samples. The sharpness and resolution of the gel bands in the experimental group were comparable to those in the control group, and the batch-to-batch repeatability was better.

[0054] Cost analysis: Taking the preparation of four concentrations as an example, the cost of the experimental group was only 35% of that of the control group.

[0055] The kit of this invention was stored at 4°C for 6 months, and performance tests were conducted at 1, 3, and 6 months. The results showed that the properties of each component were stable, and the polymerization time and electrophoretic properties of the prepared gels did not change significantly, proving that the invention has good storage stability.

[0056] The kit of this invention was tested in three different laboratories by operators with varying levels of experience. The results showed that all operators were able to successfully prepare gels of multiple concentrations within 15 minutes, demonstrating the good versatility and reproducibility of this invention.

[0057] In summary, this embodiment demonstrates significant advantages in terms of speed, convenience, flexibility across multiple concentrations, and economy, making it highly valuable for widespread application. It can be widely used in protein analysis in fields such as biomedical research, clinical testing, and drug development. Its standardized and modular design facilitates large-scale production and quality control, giving it significant industrialization prospects and market competitiveness.

[0058] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-disclosed technical content to create equivalent embodiments without departing from the scope of the present invention. Any indirect modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A simple and rapid multi-concentration gel preparation kit, comprising a lower gel layer, a upper gel layer, and a coagulant, characterized in that, The upper adhesive layer includes upper adhesive liquid A and upper adhesive liquid B; The lower layer adhesive A solution includes lower layer adhesive A1 solution, lower layer adhesive A2 solution and lower layer adhesive B solution. The lower layer adhesive A1 solution and the lower layer adhesive A2 solution are mixed with ultrapure water to form lower layer adhesive A solutions of different concentrations.

2. The simple and rapid multi-concentration gel preparation kit according to claim 1, characterized in that, The lower layer adhesive A1 solution is a mixed mother liquor of Acr and Bis; The lower layer gel A2 solution includes TEMED and ultrapure water; The lower layer adhesive B solution comprises Tris-HCl and SDS; The upper layer adhesive A solution includes Acr-Bis, TEMED, and ultrapure water; The upper layer of adhesive B solution includes Tris-HCl, SDS and ultrapure water; The coagulant is ammonium persulfate.

3. The simple and rapid multi-concentration gel preparation kit according to claim 2, characterized in that, In the lower layer of adhesive A1 solution, the concentration of the mixed mother liquor is 30%, and the ratio of Acr to Bis is 29:1; In the lower layer of adhesive A2 solution, the ratio of TEMED to ultrapure water is 0.16:12; In the lower layer of gel B solution, the pH of Tris-HCl is 8.8 and the SDS content is 10%. In the upper layer of adhesive A solution, the concentration of Acr-Bis is 30%, and the ratio of Acr-Bis, TEMED and ultrapure water is 34:0.2:

66. In the upper gel B solution, the pH of Tris-HCl is 6.8, and the ratio of Tris-HCl, SDS and ultrapure water is 26:2:

72. The concentration of ammonium persulfate in the coagulant is 10%.

4. The simple and rapid multi-concentration gel preparation kit according to claim 3, characterized in that, The lower layer adhesive A solution is composed of lower layer adhesive A1 solution, lower layer adhesive A2 solution and ultrapure water mixed in the following volume ratio: 26:3.04:

21.

5. The simple and rapid multi-concentration gel preparation kit according to claim 3, characterized in that, The lower layer adhesive A solution is composed of lower layer adhesive A1 solution, lower layer adhesive A2 solution and ultrapure water in the following volume ratio: 34:3.04:

13.

6. The simple and rapid multi-concentration gel preparation kit according to claim 3, characterized in that, The lower layer adhesive A solution is composed of lower layer adhesive A1 solution, lower layer adhesive A2 solution and ultrapure water in the following volume ratio: 40:3.04:

7.

7. The simple and rapid multi-concentration gel preparation kit according to claim 3, characterized in that, The lower layer adhesive A solution is composed of lower layer adhesive A1 solution, lower layer adhesive A2 solution and ultrapure water mixed in the following volume ratio: 46:3.04:

1.

8. A simple and rapid method for preparing multi-concentration adhesives, characterized in that, This includes preparing separating gels of different concentrations using the gel preparation kit described in any one of claims 1-7.

9. A simple and rapid multi-concentration adhesive preparation method according to claim 8, characterized in that, The steps for preparing separating gels of different concentrations are as follows: Step S1: Prepare the coagulant accelerator; Step S2: Sequentially measure the lower layer gel A1 solution, lower layer gel A2 solution, and ultrapure water to prepare the lower layer gel A solution of the corresponding concentration. Step S3: Take the lower layer adhesive A liquid, the lower layer adhesive B liquid and the coagulant and mix them to obtain the lower layer adhesive; take the upper layer adhesive A liquid and the upper layer adhesive B liquid and mix them with the coagulant to obtain the upper layer liquid. Step S4, Step S4, quickly pour the mixture into the gel mold and cover it with a layer of isopropanol to seal it; Step S5: Let stand at room temperature for 8-12 minutes. Observing a clear interface indicates that polymerization is complete.

10. A simple and rapid multi-concentration adhesive preparation method according to claim 9, characterized in that, The concentration of the lower layer adhesive A solution is 8%, 10%, 12%, or 14%.

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