An electrochemical synthesis method for target products based on mobile gels

Abstract

This invention discloses an electrochemical synthesis method for a target product based on a movable gel, comprising the following steps: preparing gels containing different reactants; reacting different gels with electrodes; simultaneously cleaning the electrodes with blank gels; repeating the above steps to complete a multi-step synthesis reaction to obtain the target product. This invention utilizes gel adsorption of reagents used in different synthesis steps, confining the reagents within the gel. DNA synthesis on the electrodes is completed through sequential contact between different gels and electrodes. The reaction process does not require complex liquid manipulation, and wiping the electrodes with gel replaces liquid rinsing. This method is simple and efficient, reducing the consumption of expensive reagents, avoiding corrosion of precision electronic equipment by liquid leakage, simplifying the complex piping and valves required for liquid manipulation, and lowering the cost of DNA synthesis. It provides new ideas for the improvement of DNA synthesis technology, the field of DNA data storage, and practical applications in electrochemical synthesis.

Description

Technical Field

[0001] This invention belongs to the field of electrochemical synthesis and DNA synthesis technology, specifically relating to an electrochemical synthesis method for a target product based on a mobile gel. Background Technology

[0002] Phosphoramide-based chemical synthesis is currently the mainstream method for DNA synthesis. Existing chemical DNA synthesis technologies include first-generation synthesis (column-based oligonucleotide synthesis) and second-generation synthesis (inkjet printing, photochemical methods, and electrochemical methods). Among these, electrochemical methods are simple and easily miniaturized, making them promising for integrated DNA storage systems. However, these synthetic methods are all carried out in a liquid-phase environment, and the reactions may produce corrosive or toxic organic liquids, acids, and oxidizing agents, which can pose hazards to the environment and operators. Furthermore, liquid leaks can cause corrosion or short circuits in the delicate microelectronic equipment used for data storage. Secondly, liquid operations require additional pumps, valves, and piping, making the entire system bulky and difficult to integrate with storage devices. Additionally, due to diffusion and convection, different reagents in the piping are prone to mixing and crosstalk over time, leading to the inability to recover expensive reagents and further increasing costs. These problems make DNA synthesis a significant factor limiting the large-scale practical application of DNA data storage.

[0003] Therefore, it is necessary to develop a new method to replace liquid-phase-based DNA synthesis methods. Summary of the Invention

[0004] Purpose of the invention: The technical problem to be solved by the present invention is to provide an electrochemical synthesis method for a target product based on a mobile gel.

[0005] Technical Solution: To solve the above-mentioned technical problems, this invention provides an electrochemical synthesis method for the target product based on a mobile gel, comprising the following steps:

[0006] (1) Preparation of gels containing different reactants: different gels with different reactants adsorbed on the gels respectively;

[0007] (2) Reactions of different gels obtained in step (1) with electrodes: The different gels adsorbed with the reaction reagents are coupled with the electrodes for coupling reaction, oxidation reaction and deprotection reaction respectively; at the same time, the blank gels are used to clean the electrodes respectively;

[0008] (3) Repeat steps (1) and (2) above to complete the multi-step synthesis reaction and obtain the target product.

[0009] In step (1), a gel is prepared by reacting monomers, crosslinking agents, photoinitiators and solvents. Then, the gel is adsorbed with coupling reagents, oxidizing reagents and deprotecting reagents to obtain different gels.

[0010] Preferably, the monomer in step (1) includes N-isopropylacrylamide or acrylamide, and the crosslinking agent includes N,N-methylenebisacrylamide or polyethylene glycol diacrylate; the molecular weight of the gel monomer is preferably 20%;

[0011] Preferably, the photoinitiator in step (1) includes 2-hydroxy-2-methylpropanone or 2,2-diethoxyacetophenone;

[0012] Preferably, the solvent in step (1) includes one or both of water and dimethyl sulfoxide;

[0013] In step (1), the mixing ratio of the solvent used to prepare the gel is arbitrary. Preferably, the mass ratio of the monomer N-isopropylacrylamide and the crosslinking agent N,N-methylenebisacrylamide is 10:1.

[0014] In step (1), when the solvent is a mixed solution of dimethyl sulfoxide and water, the volume ratio of dimethyl sulfoxide to water is 3:1 to 1:3.

[0015] In step (2), the electrode is one or more of glass slide electrode, screen-printed electrode or ITO conductive glass; the shape of the electrode includes circle, square, rectangle or polygon; the diameter or diagonal length of the electrode is 0.1 to 2 mm.

[0016] Preferably, the coupling reaction step (2) includes: placing the gel that adsorbs the coupling reagent on the electrode surface, reacting at room temperature, and simultaneously setting a blank gel to absorb only acetonitrile to wipe the electrode surface.

[0017] Preferably, the oxidation reaction step (2) includes: placing the gel adsorbed with the oxidizing agent on the electrode surface, reacting at room temperature, and wiping the electrode surface with a blank gel.

[0018] Preferably, the deprotection reaction step (2) includes: placing the gel adsorbed with the deprotection reagent on the electrode surface, applying voltage, and wiping the electrode surface with blank gel after the reaction; selectively deprotecting the microelectrode array surface, applying voltage to the electrode that needs to be used for the next step of base synthesis, reacting, and finally wiping with blank gel.

[0019] In step (2), the electrolyte in the electrochemical reaction of the present invention includes substances capable of electrolyzing hydrogen ions at the anode, including hydroquinone and tetraethylamine perchlorate; the electrolyte includes substances capable of reducing hydrogen ions at the cathode, including benzoquinone and anthraquinone; the electrolyte contains stable electrolytes, including tetraethyl toluenesulfonate and tetrabutylammonium hexafluorophosphate; and the electrolyte contains stable solvents, such as acetonitrile, dichloromethane, methanol, and mixtures thereof. Preferably, the electrolyte in which the gel is immersed includes hydroquinone, benzoquinone, and tetrabutylammonium hexafluorophosphate, and the solvent is acetonitrile.

[0020] In step (3), the solvent adsorbed by the blank gel used to clean the electrode can be any reagent that does not affect the normal electrochemical reaction. Preferably, anhydrous acetonitrile is used.

[0021] Preferably, the electrochemical reaction applied voltage of the present invention is in the range of 1.2V-1.8V, and more preferably, 1.4V is used.

[0022] Furthermore, the specific application of the electrochemical synthesis can be polymers composed of a limited number of monomers, such as DNA, RNA, and polypeptides.

[0023] Further, in step (2), the steps of using the gel on the electrode surface for DNA synthesis include DNA coupling, oxidation, deprotection and washing; the DNA coupling time of the gel on the electrode surface is 2 minutes, the DNA oxidation time of the gel on the electrode surface is 40 seconds, and the DNA deprotection time of the gel on the electrode surface is 14 seconds.

[0024] In step (2), the coupling reagent includes different phosphoramide monomers (DMT-dA(bz) phosphoramide monomer, DMT-dG(ib) phosphoramide monomer, DMT-dC(bz) phosphoramide monomer, DMT-d phosphoramide monomer) and activator 5-ethylthiotetrazole.

[0025] In step (2), the oxidizing agent includes trichloroperoxybenzoic acid.

[0026] In step (2), the deprotection reagent includes hydroquinone, p-benzoquinone, and tetrabutylammonium hexafluorophosphate.

[0027] In step (3), the above steps (1) and (2) are repeated until all bases on the electrode are synthesized.

[0028] Preferably, the base sequence is as shown in SEQ ID NO: 1.

[0029] The present invention also includes the target product obtained by the electrochemical synthesis method, wherein the target product may be a polymer composed of a limited number of monomers, such as DNA, RNA, or polypeptide.

[0030] Furthermore, in step (2), the surface of the microelectrode array is modified before the reaction:

[0031] (a) Surface cleaning: Immerse in piranha solution (volume ratio of concentrated sulfuric acid: hydrogen peroxide = 1:1) for 15 min, rinse with ultrapure water, and then perform ultrasonic cleaning with alternating acetone, ethanol, and water;

[0032] (b) Silanization: The cleaned electrode and a bottle containing 2% 1H,1H,2H,2H-perfluorodecyltriethoxysilane dissolved in 2 mL of dichloromethane were placed in the same sealed container and heated at 80°C for more than 2 hours, and then cooled to room temperature.

[0033] (c) Modification with mercaptohexanol: 20 mM 6-mercapto-1-hexanol was added dropwise to the surface of the working electrode and incubated at room temperature for 12 h;

[0034] Furthermore, in step (2), DNA synthesis is performed on the electrode surface:

[0035] (a) Coupling reaction: The gel adsorbed with coupling reagent (0.1M monomer and 0.5M activator 5-ethylthiotetrazole) was placed on the electrode surface and reacted at room temperature for 2 min. The electrode surface was wiped with blank gel (which only absorbs acetonitrile).

[0036] (b) Oxidation reaction: The gel that adsorbs the oxidizing agent (0.1M trichloroperoxybenzoic acid) is placed on the electrode surface and reacted at room temperature for 40 seconds. The electrode surface is then wiped with a blank gel (which only absorbs acetonitrile).

[0037] (c) Deprotection reaction: A gel containing the deprotection reagents (25 mM hydroquinone, 25 mM p-benzoquinone and 25 mM tetrabutylamine hexafluorophosphate) was placed on the electrode surface and reacted for 14 seconds with an electric field of 1.4 V. The electrode surface was then wiped with a blank gel (which only absorbs acetonitrile).

[0038] (d) Repeat steps (a) to (c) above until all bases on the electrode are synthesized.

[0039] This invention utilizes gel adsorption to absorb reagents used in different synthesis steps (DNA coupling, oxidation, deprotection, and washing). The reagents are confined within the gel, and DNA synthesis on the electrodes is completed through sequential contact between different gels and electrodes. The reaction process does not require complex liquid manipulation, and wiping the electrodes with gel replaces liquid rinsing. This method is simple and efficient, reducing the consumption of expensive reagents, avoiding corrosion of precision electronic equipment by liquid leakage, simplifying the complex piping and valves required for liquid manipulation, and reducing the cost of DNA synthesis. It provides new ideas for the improvement of DNA synthesis technology, the practical application of DNA data storage, and electrochemical synthesis.

[0040] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0041] (1) This invention proposes an electrochemical synthesis method for target products based on mobile gels. By utilizing the solid-liquid properties of gels, the reaction solvent can be fully absorbed and kept in a saturated state, while maintaining a certain mechanical strength and allowing various reactants to diffuse to the reaction interface, thus eliminating the need for complex liquid manipulation steps.

[0042] (2) The method of the present invention utilizes gel adsorption of reagents remaining in the reaction area, eliminating the need for liquid cleaning steps;

[0043] (3) The method of the present invention is based on electrochemical synthesis reaction, and can directly use commercial reagents for the synthesis of DNA, RNA and polypeptides, which has a wide range of applications. Attached Figure Description

[0044] Figure 1 This is a schematic diagram of the electrochemical synthesis method of the target product based on a mobile gel according to the present invention;

[0045] Figure 2 This is a graph showing the relationship between the relative mass and time of gels prepared using different solvents according to the present invention;

[0046] Figure 3 This is a photograph of the N-isopropylacrylamide (NIPAM) gel involved in this invention.

[0047] Figure 4 This is a schematic diagram of the microelectrode array structure involved in the present invention;

[0048] Figure 5 Raman spectra before and after wiping the electrode surface with blank gel;

[0049] Figure 6 The fluorescence image shows the hybridization of DNA synthesized on the electrode surface with complementary fluorescent chains.

[0050] Figure 7 The relationship between the amount of DNA synthesized on the electrode surface and the square root of time. Detailed Implementation

[0051] The technical solution of the present invention will be further described below with reference to the accompanying drawings.

[0052] like Figure 1 The present invention provides an electrochemical synthesis method for a target product based on a mobile gel, comprising the following steps:

[0053] (1) Transfer the reaction gel to the electrode surface;

[0054] Specifically, the saturated gel containing the adsorbed reagents is transferred to the surface of the reaction electrode.

[0055] The NIPAM gel was completely dried by high-temperature heating, and then immersed in the reaction reagent until saturation. The molecular weight of the gel monomer was 20 wt%.

[0056] (2) Take the saturated gel obtained in step (1) onto the electrode surface, and the internal reaction reagent gradually diffuses to the electrode surface and reacts with the electrode surface substance;

[0057] The synthesis reaction steps include: coupling reaction, oxidation reaction, and deprotection reaction. The above steps are repeated until all bases on the electrode are synthesized. The reaction steps are wiped with blank gel between steps.

[0058] (3) Remove the gel and blank gel used for wiping from the electrode surface after each reaction in step (2).

[0059] Specifically, the gel after the reaction and the blank gel after wiping are removed from the electrode surface.

[0060] Example 1: Optimization of Gel Preparation Conditions

[0061] The steps for optimizing the solvent ratio used in the gel are as follows:

[0062] (1) Five different gel solvent ratios were designed, namely, the ratio of dimethyl sulfoxide to deionized water was 0:1, 1:3, 1:1, 3:1 and 1:0.

[0063] Specifically, it includes the following five types:

[0064] Weigh 0.6g of monomer N-isopropylacrylamide, 0.06g of N,N-methylenebisacrylamide, and 3mL of deionized water, vortex at room temperature until completely dissolved to obtain a gel prepolymer solution;

[0065] Weigh 0.6g of monomer N-isopropylacrylamide, 0.06g of N,N-methylenebisacrylamide, 0.75mL of dimethyl sulfoxide and 2.25mL of deionized water, vortex at room temperature until completely dissolved to obtain a gel prepolymer solution;

[0066] Weigh 0.6g of monomer N-isopropylacrylamide, 0.06g of N,N-methylenebisacrylamide, 1.5mL of dimethyl sulfoxide and 1.5mL of deionized water, vortex at room temperature until completely dissolved to obtain a gel prepolymer solution;

[0067] Weigh 0.6g of monomer N-isopropylacrylamide, 0.06g of N,N-methylenebisacrylamide, 2.25mL of dimethyl sulfoxide and 0.75mL of deionized water, vortex at room temperature until completely dissolved to obtain a gel prepolymer solution;

[0068] Weigh 0.6g of monomer N-isopropylacrylamide, 0.06g of N,N-methylenebisacrylamide, and 3mL of dimethyl sulfoxide. Vortex at room temperature until completely dissolved to obtain a gel prepolymer solution.

[0069] (2) Add 30 μL of 2-hydroxy-2-methylpropanone to the five gel prepolymer solutions obtained in (1) above, irradiate with a UV lamp (5W) for 45 seconds to obtain the corresponding gels, place them on a hot table or oven until completely dry, record the mass as m0, and soak them in anhydrous acetonitrile.

[0070] (3) Every minute, remove the gel from the acetonitrile and record its mass as m. t Record continuously for more than ten minutes.

[0071] (4) Calculate the relative mass (m) of the gel after different times. t / m0), and calculate the relative mass versus time as follows: Figure 2 As shown, when the ratio of dimethyl sulfoxide to deionized water is 3:1, the gel absorbs acetonitrile and swells the fastest.

[0072] Example 2: Gel Preparation

[0073] The preparation steps of the gel used in this invention are as follows:

[0074] (1) Weigh 0.6g of monomer N-isopropylacrylamide, 0.06g of N,N-methylenebisacrylamide, 0.75mL of deionized water and 2.25mL of dimethyl sulfoxide, and vortex at room temperature until completely dissolved to obtain a gel prepolymer solution.

[0075] (2) Add 30 μL of 2-hydroxy-2-methylpropanone to the solution obtained in step (1) above, and vortex at room temperature until completely dissolved.

[0076] (3) Using a pipette, take the solution obtained in step (2) above and add it between two glass plates, with a thickness of 1.2 mm between the two glass plates. Irradiate with a 5W UV lamp for 45 seconds to obtain N-isopropylacrylamide gel, as shown. Figure 3 As shown. Soak the gel in deionized water overnight.

[0077] (4) Take the gel obtained in step (3) above onto the surface of a glass slide, heat it on a hot table or oven at 80°C for more than 30 minutes to obtain a completely dry gel.

[0078] Example 3: Fabrication of a microelectrode array and modification of its surface.

[0079] 1. The steps for fabricating the microelectrode array are as follows:

[0080] (1) Design the microelectrode array pattern and commission a company to process the mask. The circular electrodes have a diameter of 1 mm and a spacing of 1 mm, while the square antennae have a side length of 1 mm and a spacing of 1.27 mm. The designed microelectrode array pattern is as follows: Figure 4 .

[0081] (2) Photolithography steps: Clean the glass slide, homogenize the coating, pre-bake, expose, and develop in sequence.

[0082] (3) Evaporation steps: Place the glass slide obtained in step (2) above into the evaporation chamber and perform vacuuming, preheating the target material, evaporating the adhesive layer titanium metal, cooling the target material, preheating the target material, and evaporating the electrode layer gold metal in sequence.

[0083] (4) Immerse the glass slide obtained in step (3) above in the stripping solution, incubate at room temperature for more than 4 hours, rinse thoroughly with ethanol and water, and dry in an oven at 60°C for two hours.

[0084] 2. Modify the surface of the microelectrode array before the reaction:

[0085] (a) Surface cleaning: Immerse in piranha solution (volume ratio of concentrated sulfuric acid: hydrogen peroxide = 1:1) for 15 min, rinse with ultrapure water, and then perform ultrasonic cleaning with alternating acetone, ethanol, and water;

[0086] (b) Silanization: The cleaned electrode and a bottle containing 2% 1H,1H,2H,2H-perfluorodecyltriethoxysilane dissolved in 2 mL of dichloromethane were placed in the same sealed container and heated at 80°C for more than 2 hours, and then cooled to room temperature.

[0087] (c) Mercaptohexanol modification: 20 mM 6-mercapto-1-hexanol was dropped onto the surface of the working electrode and incubated at room temperature for 12 h. Example 4: Characterization of the effect of blank gel wiping the electrode.

[0088] (1) The bare gold electrode surface was used as a control group, and its surface Raman spectrum was tested. The excitation wavelength was 632 nm, the laser power was 5 mW, and the integration time was 10 seconds.

[0089] (2) The completely dry gel prepared in Example 2 was immersed in acetonitrile dissolved in cy3-modified phosphoramidite monomer (Hebei Dina Xingke Biotechnology Co., Ltd.-20220310). After saturation, it was taken out, placed on the electrode surface, and then removed to test its surface Raman spectrum with the same parameters as in step (1).

[0090] (3) Transfer the blank gel to the reaction area and wipe the electrode from left to right in one direction for 10 seconds. Measure the surface Raman spectrum after wiping, using the same parameters as in step (1). The final Raman spectra of the blank gel before and after wiping the electrode are shown below. Figure 5 As shown.

[0091] Example 5: Electrochemical DNA Synthesis Based on Different Gels

[0092] Different reagents used in DNA synthesis were adsorbed onto different gels until saturation was achieved, and the process was repeated sequentially.

[0093] (1) Coupling reaction: A gel containing 0.1M monomers (including DMT-dA(bz) phosphorus amide monomer, DMT-dG(ib) phosphorus amide monomer, DMT-dC(bz) phosphorus amide monomer, and DMT-d phosphorus amide monomer, all at a concentration of 0.1M) and 0.5M activator 5-ethylthiotetrazole) was placed on the electrode surface and reacted at room temperature for 2 minutes. The electrode surface was then wiped with a blank gel (which only absorbs acetonitrile).

[0094] (2) Oxidation reaction: The gel adsorbed with oxidizing agent (0.1M trichloroperoxybenzoic acid) was placed on the electrode surface and reacted at room temperature for 40 seconds. The electrode surface was wiped with blank gel.

[0095] (3) Deprotection reaction: The gel adsorbed with deprotection reagents (25mM hydroquinone, 25mM p-benzoquinone and 25mM tetrabutylamine hexafluorophosphate) was placed on the electrode surface, a voltage of 1.4V was applied, and the reaction was carried out for 14 seconds. The electrode surface was then wiped with a blank gel. Selective deprotection can be performed on the surface of the microelectrode array. The electrode that needs to be used for the next step of base synthesis was selected, a voltage of 1.4V was applied, the reaction time was 14 seconds, and finally it was wiped with a blank gel.

[0096] (4) Repeat (1) to (3) above until all bases on the electrode are synthesized (5'-TTGTCTGTATGTTTTTT-3');

[0097] (5) The electrode was immersed in a mixed solution of ethylenediamine and ethanol at a volume ratio of 1:1. After removing the side-chain protecting groups at room temperature for two hours, it was hybridized with 25 μM complementary fluorescent single-stranded electron (5'-ACATACAGACAA-Cy3-3'). The reaction was carried out in the dark for 1 hour, and the results were observed under a fluorescence microscope. Figure 6 As shown, electrode (A) that performed the DNA synthesis experiment hybridized with the fluorescent chain and showed obvious fluorescence, while the control group (B) was a blank electrode (i.e., no DNA synthesis experiment was performed), which was clearly distinguishable from the experimental group.

[0098] Example 6: Characterization of DNA synthesized on the electrode surface

[0099] The characterization steps for the DNA synthesized on the electrode surface are as follows:

[0100] (1) Weigh 0.012 g of hexaammineruthenium trichloride, dissolve it in 10 mM Tris-HCl (pH 8.3) to a final concentration of 20 mM and a volume of 2 mL, and further dilute it to 50 μM.

[0101] (2) Puff nitrogen gas into the Tris-HCl buffer (10mM, pH 8.3) and the 50μM hexaammineruthenium trichloride solution obtained in step (1) for 10 minutes.

[0102] (3) Take 50 μL of the deoxygenated Tris-HCl buffer solution obtained in step (2) and 50 μM hexaammineruthenium trichloride solution and add them dropwise to the electrode surface (polyT-12 has been synthesized). Measure using a chronocoulometric method. The starting voltage is 0.2 V, the ending voltage is -0.5 V, the number of steps is 2, the pulse duration is 0.25 s, and the measurement sensitivity is 1 × 10⁻⁶. -5 A / V.

[0103] (4) Plot the measurement results obtained in step (3) above as a curve of charge versus the square root of time, fit the curve to a straight line and obtain the intercept difference, i.e., the charge difference, as shown below. Figure 7 As shown, there is a significant charge difference on the electrode surface (A) where DNA is synthesized, with a difference of about 30 nanocoulombs, while there is no charge difference on the bare gold electrode surface (B).

Claims

1. An electrochemical synthesis method for a target product based on a mobile gel, characterized in that, Includes the following steps: (1) Preparation of gels containing different reactants: different gels with different reactants adsorbed on the gels respectively; (2) React the different gels obtained in step (1) with the electrodes: The different gels adsorbed with the reaction reagents are coupled with the electrodes for coupling reaction, oxidation reaction and deprotection reaction respectively; at the same time, the blank gels are used to clean the electrodes respectively; The coupling reaction step includes: placing a gel adsorbing the coupling reagent on the electrode surface, reacting at room temperature, and simultaneously setting a blank gel to absorb only acetonitrile to wipe the electrode surface; the oxidation reaction step includes: placing a gel adsorbing the oxidizing reagent on the electrode surface, reacting at room temperature, and wiping the electrode surface with a blank gel; the deprotection reaction step includes: placing a gel adsorbing the deprotection reagent on the electrode surface, applying voltage, reacting, and then wiping the electrode surface with a blank gel; selective deprotection is performed on the microelectrode array surface, selecting the electrode for the next step of base synthesis, applying voltage, reacting, and finally wiping with a blank gel; (3) Repeat steps (1) and (2) above to complete the multi-step synthesis reaction and obtain the target product.

2. The electrochemical synthesis method for the target product based on a mobile gel according to claim 1, characterized in that, In step (1), a gel is prepared by reacting monomers, crosslinking agents, photoinitiators and solvents. Then, the gel is adsorbed with coupling reagents, oxidizing reagents and deprotecting reagents to obtain different gels.

3. The electrochemical synthesis method for the target product based on a mobile gel according to claim 2, characterized in that, The monomer includes N-isopropylacrylamide or acrylamide, and the crosslinking agent includes N,N-methylenebisacrylamide or polyethylene glycol diacrylate.

4. The electrochemical synthesis method for the target product based on a mobile gel according to claim 2, characterized in that, The photoinitiator includes 2-hydroxy-2-methylpropanone or 2,2-diethoxyacetophenone.

5. The electrochemical synthesis method for the target product based on a mobile gel according to claim 2, characterized in that, The solvent includes one or both of water and dimethyl sulfoxide.

6. The electrochemical synthesis method for the target product based on a mobile gel according to claim 2, characterized in that, The mass ratio of the monomer N-isopropylacrylamide to the crosslinking agent N,N-methylenebisacrylamide is 10:1 to 5:

1.

7. The electrochemical synthesis method for the target product based on a mobile gel according to claim 1, characterized in that, In step (1), when the solvent is a mixed solution of dimethyl sulfoxide and water, the volume ratio of dimethyl sulfoxide to water is 3:1 to 1:

3.

8. The electrochemical synthesis method for the target product based on a mobile gel according to claim 1, characterized in that, The electrode described in step (2) includes one or more of glass slide electrodes, screen-printed electrodes, or ITO conductive glass; the shape of the electrode includes circular, square, rectangular, or polygonal; the diameter or diagonal length of the electrode is 0.1 to 2 mm.

9. The electrochemical synthesis method for the target product based on a mobile gel according to claim 1, characterized in that, The coupling reagents include DMT-dA(bz) phosphorus amide monomer, DMT-dG(ib) phosphorus amide monomer, DMT-dC(bz) phosphorus amide monomer, DMT-d phosphorus amide monomer and activator 5-ethylthiotetrazole.

10. The electrochemical synthesis method for the target product based on a mobile gel according to claim 1, characterized in that, The oxidizing agent includes trichloroperoxybenzoic acid.

11. The electrochemical synthesis method of the target product based on a mobile gel according to claim 1, characterized in that, The deprotection agents include hydroquinone, p-benzoquinone, and tetrabutylamine hexafluorophosphate.

12. The electrochemical synthesis method for the target product based on a mobile gel according to claim 1, characterized in that, Repeat steps (1) and (2) above until all bases of the sequence SEQ ID NO: 1 on the electrode have been synthesized.

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