A preparation method of bionic nano-microstructure and application of bionic nano-microstructure in DNA synthesis

Abstract

The present invention discloses a preparation method of a biomimetic nano-microstructure and an application of the biomimetic nano-microstructure in DNA synthesis. The biomimetic nano-microstructure of the present invention is an inverse opal structure. The preparation method thereof is to scrape a coating that can form an opal structure on the surface of a substrate containing pyramidal depressions, and then form an inverse opal nano-microstructure with a higher porosity after curing the coating, dissolving the opal structure, and sintering. The process of synthesizing DNA is: preparing an inverse opal structure in the pyramidal depression of the substrate, and sequentially adding a universal solid phase carrier, a deprotecting agent, an ATCG base monomer, an activator, a capping agent, and an oxidant into the pyramidal depression by scraping. The beneficial effect of the present invention is: because the inverse opal structure has the function of selectively reflecting specific wavelengths and transmitting other wavelengths, the above-mentioned reaction steps can effectively increase the synthesis speed of DNA under the assistance of light of a specific wavelength.

Description

Technical Field

[0001] The invention relates to the field of materials and biology, and in particular to a method for preparing a bionic nano-microstructure and application of the bionic nano-microstructure in DNA synthesis. Background Art

[0002] In 1987, S. John et al. found that materials with different dielectric constants and their spatially periodic structures can produce photonic band gaps, thereby controlling the behavior of light. They proposed the concept of photonic crystals. Photonic crystals or inverse opal structures derived from photonic crystals can be used to regulate the propagation of light, thereby achieving control over chemical reactions that respond to light.

[0003] The three-dimensional pore structure of inverse opal is a structure with periodic changes in the refractive index, and its periodic scale matches the wavelength of light. When light propagates in the inverse opal structure membrane system with periodic changes in the refractive index, Bragg diffraction occurs, forming selective reflection of specific wavelengths and transmission of other wavelengths. This structure can be used for biological detection and catalyst preparation (CN 101339135B, CN 112062157A).

[0004] The method of DNA synthesis has undergone three generations of changes:

[0005] The first generation of synthesis methods is column synthesis, including phosphodiester method, phosphotriester method, phosphite triester method, phosphite amide method and solid phase phosphite amide method. This type of method uses a synthesis column filled with porous glass or polystyrene sieve plates as a solid phase carrier. Each DNA is synthesized in a separate synthesis column. The yield is high, generally in the nmol range, but the amount of reagents consumed in the synthesis is large, the cost is also high, and the synthesis throughput is limited.

[0006] The second generation of synthesis is chip synthesis. The chip is used as a solid phase carrier for DNA synthesis. The synthesis reaction is carried out at specific sites on its surface in a high-density and integrated manner, saving reagents while achieving high-throughput synthesis of DNA. The chip DNA synthesis technology is based on the four-step cycle reaction of the phosphoamidite synthesis method. Because of the use of different "deprotection" fixed-point control methods, photochemical, electrochemical, microfluidic and inkjet printing DNA synthesis methods have been developed. Among them, the photochemical synthesis method is characterized by high selectivity and controllability, but it is limited to a very small number of application scenarios due to the high cost of photosensitive deprotection monomers and the short synthesis length; the electrochemical synthesis method can achieve local high-precision DNA synthesis, but because this method needs to be carried out on a specific electrode surface and has a high synthesis error rate, it is gradually eliminated. The synthesis method based on microfluidics realizes the automation and efficiency of DNA synthesis, but the need to introduce micropumps and microvalves makes its synthesis device structure relatively complex, cumbersome to operate, and reduces efficiency, and it has not become a mainstream method. The inkjet printing DNA synthesis method has high throughput, low synthesis cost, and longer synthesis length, and has gradually become the mainstream high-throughput oligonucleotide synthesis method. However, when using inkjet printing alone, the selected material is restricted by viscosity, surface tension, etc., so the scope of application of this method is limited.

[0007] In summary, in order to solve the problems existing in the preparation methods of the above prior arts and to construct non-interfering reaction chambers on the surface of a substrate, the present invention provides a preparation method of a bionic nano-microstructure and an application of the bionic nano-microstructure in DNA synthesis. The method first prepares an inverse opal structure in a pyramidal depression of a substrate, and then sequentially scrapes a universal solid phase carrier (DMT-protected phosphite amide solid powder and an insoluble polymer, the insoluble polymer is selected from one of silica gel, cross-linked polystyrene or porous glass beads), a deprotecting agent, an ATCG base monomer, an activator, a capping agent, and an oxidant into the pyramidal depression, thereby realizing high-throughput DNA synthesis in a simple and efficient manner. Summary of the invention

[0008] The purpose of the present invention is to provide a method for preparing a bionic nano-microstructure and the application of the bionic nano-microstructure in DNA synthesis. An array of pyramidal depressions is constructed on the surface of a substrate, and then a bionic nano-microstructure is prepared in the depressions. A universal solid phase carrier, a deprotecting agent, an ATCG base monomer, an activator, a capping agent, and an oxidant required for DNA synthesis are scraped and coated, and DNA is synthesized simply and efficiently with the assistance of light of a characteristic wavelength.

[0009] To achieve the above object, the present invention is implemented by the following technical solutions:

[0010] A method for preparing a bionic nano-microstructure, the bionic nano-microstructure comprising the following preparation steps:

[0011] 1) Substrate and surface pyramidal microstructure

[0012] An array of pyramidal depressions is constructed on the surface of the substrate by photolithography;

[0013] 2) Preparation of inverse opal biomimetic nanostructures

[0014] Prepare biomimetic nanostructures in the depressions.

[0015] As a further improvement of the present scheme, the bionic nanostructure is an inverse opal structure, and its preparation process is as follows: first, a coating that can form an opal structure is scraped on the surface of the cleaned substrate, and then the coating is cured, the opal structure is dissolved, and sintered to form an inverse opal nanostructure with a higher porosity.

[0016] As a further improvement of this scheme,

[0017] The substrate is any one of a silicon substrate, a silicon dioxide substrate, a quartz substrate or a glass substrate;

[0018] The array on the surface of the substrate is composed of pyramidal depressions with an upper side length of 5-1000 μm, an edge depression thickness of 1-20 μm, a bottom side length of 1-500 μm, and a height of 0.05-50 μm.

[0019] The angle of the pyramid ranges from 0.5 to 90 degrees, and there are micropores at the bottom of the depression with a pore size of 1 to 100 μm.

[0020] The cleaning is to ultrasonically clean the substrate in sequence using ethanol, acetone, and isopropanol.

[0021] As a further improvement of this solution, the coating is composed of organic or inorganic microspheres, a supporting material and a solvent.

[0022] As a further improvement of the present invention, the organic microspheres are composed of any one of polystyrene, styrene-acrylic acid copolymer, styrene-methyl acrylate copolymer, styrene-butyl acrylate copolymer, styrene-methacrylic acid copolymer, styrene-methyl methacrylate copolymer, styrene-butyl methacrylate copolymer, styrene-acrylic acid-methyl acrylate copolymer, styrene-acrylic acid-methyl methacrylate copolymer, styrene-acrylic acid-butyl acrylate copolymer, and styrene-acrylic acid-butyl methacrylate copolymer;

[0023] The inorganic microsphere composition is selected from any one of silicon dioxide and titanium dioxide.

[0024] As a further improvement of this solution, the support material is selected from any one or more of amorphous carbon, phenolic resin, epoxy resin, polystyrene, polyurethane, epoxy-acrylic resin, polyurethane acrylic resin, and phenolic epoxy acrylic resin;

[0025] The mass composition ratio of the organic or inorganic microspheres to the supporting material in the coating is 1:5-5:1.

[0026] As a further improvement of this solution, the coating solvent is selected from any one or more of water, ethanol, ethyl acetate, 6-caprolactone, butyl acetate, ethylene glycol methyl ether, ethylene glycol butyl ether, ethylene glycol phenyl ether, and propylene glycol methyl ether acetate;

[0027] The curing of the substrate surface coating is thermal curing, the heating temperature is 80-150° C., and the curing time is 5-15 minutes; the dissolution is to dissolve the organic or inorganic microspheres with toluene or concentrated sulfuric acid;

[0028] The coating sintering temperature is 100-250°C and the sintering time is 10-20 minutes.

[0029] As a further improvement of this solution, the diameter of the inverse opal structure ranges from 500 to 1000 nm.

[0030] An application of a bionic nano-microstructure in DNA synthesis, wherein the bionic nano-microstructure is prepared by a bionic nano-microstructure preparation method;

[0031] DNA synthesis, including the following steps:

[0032] (1) Fill the inverse opal structure with a universal solid phase carrier by scraping and sintering at 120-200°C for 5-10 minutes.

[0033] (2) After cooling to room temperature, the deprotecting agent, ATCG base monomer, activator, capping agent, and oxidant are sequentially scraped into the inverse opal structure to synthesize DNA. The characteristic wavelength of the auxiliary light for each reaction is 350-2000nm (matching the inverse opal structure), and washing is required after the reaction.

[0034] As a further improvement of this scheme,

[0035] The deprotecting agent is a trichloroacetic acid-dichloromethane solution, the solution composition is 5-15wt% trichloroacetic acid and the balance dichloromethane;

[0036] The activator is a 5-ethylmercaptotetrazole-acetonitrile solution, the solution composition is 5-15wt% 5-ethylmercaptotetrazole and the balance acetonitrile;

[0037] The capping agent is acetic anhydride-tetrahydrofuran solution and N-methylimidazole-tetrahydrofuran solution, and the solution composition is 10-25wt% of acetic anhydride, 5-10wt% of N-methylimidazole and the balance of tetrahydrofuran;

[0038] The oxidant is a solution of iodine-pyridine-tetrahydrofuran-water, wherein the oxidant solution contains 5-10wt% iodine, 10-15wt% pyridine, 40-60wt% tetrahydrofuran and the balance is water.

[0039] As a further improvement of this scheme, the specific reaction steps of the DNA synthesis are:

[0040] 1) Fill the inverse opal structure with a universal solid phase carrier by scraping and sintering after filling;

[0041] 2) After the substrate is cooled to room temperature, apply the deprotection agent solution, and after sufficient reaction, rinse with a large amount of acetonitrile to remove the unreacted reactants;

[0042] 3) Apply acetonitrile solution and activator solution containing 1-20wt% ATCG base monomer in sequence, and rinse with a large amount of acetonitrile after sufficient reaction to remove unreacted reactants;

[0043] 4) Scrape the capping agent solution and rinse with a large amount of acetonitrile after sufficient reaction to remove the unreacted reactants;

[0044] 5) Scrape the oxidant solution and rinse with a large amount of acetonitrile after sufficient reaction to remove the unreacted reactants.

[0045] As a further improvement of the present solution, the characteristic wavelength of the auxiliary light in each step of the reaction is 350-2000nm. Preferably, the characteristic wavelength of the auxiliary light is 350-600nm, 600-1500nm or 1500-1800nm.

[0046] The preparation method of a bionic nano-microstructure and the application of the bionic nano-microstructure in DNA synthesis of the present invention have the following beneficial effects:

[0047] (1) The present invention is the first to use an inverse opal bionic nanostructure and a spraying method to combine with synthetic DNA.

[0048] (2) The present invention designs a special pyramidal recessed structure array on the surface of the substrate.

[0049] (3) The present invention utilizes the characteristic that light of a specific wavelength has a photon bandgap in the inverse opal structure.

[0050] (4) In the present invention, since the inverse opal structure has the function of selectively reflecting specific wavelengths and transmitting other wavelengths, the above-mentioned reaction steps can effectively increase the synthesis rate of DNA under the assistance of light of specific wavelengths.

[0051] (5) The method of the present invention is characterized by low cost, simplicity, high efficiency and high throughput.

[0052] (6) The bionic nanostructure of the present invention can be further applied to other biosynthesis fields that require nanostructures. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1 It is one of the inverse opal bionic nanostructures used in the present invention.

[0054] Figure 2 Schematic diagram of the relationship between the substrate morphology and the position of the inverse opal structure designed for the present invention. DETAILED DESCRIPTION

[0055] In order to make the purpose, technical solutions and advantages of the present invention more clear, the following embodiments and the accompanying drawings are combined to illustrate the present invention. Figure 1~2 The present invention will be further described:

[0056] Example 1

[0057] Substrate and surface pyramidal microstructure:

[0058] The substrate is a silicon substrate, and the surface is composed of a pyramidal recessed structure array with an upper side length of 5 μm, a recessed edge thickness of 1 μm, a bottom side length of 1 μm, and a height of 0.05 μm. The angle of the pyramid is 0.5 degrees, and there are micropores at the bottom of the recess with a pore diameter of 1 μm. The substrate is ultrasonically cleaned in sequence with ethanol-acetone-isopropanol.

[0059] Preparation of inverse opal biomimetic nanostructures:

[0060] A solution consisting of 0.2wt% styrene-acrylic copolymer microspheres, 1wt% phenolic resin and the remainder of ethylene glycol methyl ether was scraped on the surface of the cleaned substrate and cured at 150°C for 5 min. The microspheres were dissolved in toluene and then sintered at 250°C for 10 min. The diameter of the prepared inverse opal bionic nanostructure was 500-550nm.

[0061] Synthetic DNA:

[0062] 1) Scrape a universal solid phase carrier on the surface of the substrate and sinter at 120°C for 10 minutes; 2) After the substrate is cooled to room temperature, scrape a 5wt% trichloroacetic acid-dichloromethane solution, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants; 3) Scrape ATCG base monomer 1wt% acetonitrile solution and 5wt% 5-ethylmercaptotetrazole-acetonitrile solution alternately, and rinse with a large amount of acetonitrile after sufficient reaction to remove the unreacted reactants; 4) Scrape 10wt% acetic anhydride-tetrahydrofuran solution and 5wt% N-methylimidazole-tetrahydrofuran solution, and rinse with a large amount of acetonitrile after sufficient reaction to remove the unreacted reactants; 5) Scrape an oxidant solution with an iodine content of 5wt%, pyridine 15wt%, tetrahydrofuran 60wt% and the remainder of water, and rinse with a large amount of acetonitrile after sufficient reaction to remove the unreacted reactants. The characteristic wavelength of the auxiliary light in the above steps is 1000-1100nm.

[0063] Example 2

[0064] Substrate and surface pyramidal microstructure:

[0065] The substrate is a silicon dioxide substrate, and the surface is composed of a pyramidal recessed structure array with an upper side length of 1000 μm, a bottom side length of 500 μm, a recessed edge thickness of 20 μm, and a height of 50 μm. The angle of the pyramid is 90 degrees, and there are micropores at the bottom of the recess with a pore diameter of 100 μm. The substrate is ultrasonically cleaned in sequence with ethanol-acetone-isopropanol.

[0066] Preparation of inverse opal biomimetic nanostructures:

[0067] A solution consisting of 1wt% silica microspheres, 0.2wt% polyurethane and the remainder ethylene glycol butyl ether was coated on the surface of the cleaned substrate and cured at 80°C for 15 min; the microspheres were dissolved with concentrated sulfuric acid and then sintered at 100°C for 20 min; the diameter of the prepared inverse opal bionic nanostructure was 950-1000nm.

[0068] Synthetic DNA:

[0069] 1) Scrape a universal solid phase carrier on the surface of the substrate and sinter at 200°C for 5 minutes; 2) After the substrate is cooled to room temperature, scrape a 15wt% trichloroacetic acid-dichloromethane solution, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants; 3) Scrape 20wt% acetonitrile solution of ATCG base monomer and 15wt% 5-ethylthiotetrazolyl-acetonitrile solution alternately, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants; 4) Scrape 25wt% acetic anhydride-tetrahydrofuran solution and 5wt% N-methylimidazole-tetrahydrofuran solution, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants; 5) Scrape an oxidant solution with an iodine content of 10wt%, pyridine 15wt%, tetrahydrofuran 60wt% and the remainder of water, and rinse with a large amount of acetonitrile after sufficient reaction to remove the unreacted reactants. The characteristic wavelength of the auxiliary light in the above steps is 1900-2000nm.

[0070] Example 3

[0071] Substrate and surface pyramidal microstructure:

[0072] The substrate is a silicon substrate, and the surface is composed of a pyramidal recessed structure array with an upper side length of 100 μm, a recessed edge thickness of 10 μm, a bottom side length of 20 μm, and a height of 10 μm. The angle of the pyramid is 60 degrees, and there are micropores at the bottom of the recess with a pore diameter of 10 μm. The substrate is ultrasonically cleaned in sequence with ethanol-acetone-isopropanol.

[0073] Preparation of inverse opal biomimetic nanostructures:

[0074] A solution consisting of 0.2wt% polystyrene microspheres, 0.3wt% epoxy resin and the remainder of ethanol was coated on the surface of the cleaned substrate and cured at 100°C for 5 min; the microspheres were dissolved in toluene and then sintered at 150°C for 15 min. The diameter of the prepared inverse opal bionic nanostructure was 600-650nm.

[0075] Synthetic DNA:

[0076] 1) Scrape a universal solid phase carrier on the surface of the substrate and sinter at 150°C for 8 minutes; 2) After the substrate is cooled to room temperature, scrape a 10wt% trichloroacetic acid-dichloromethane solution, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants; 3) Scrape alternately a 10wt% acetonitrile solution of ATCG base monomer and a 10wt% 5-ethylmercaptotetrazole-acetonitrile solution, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants; 4) Scrape a 15wt% acetic anhydride-tetrahydrofuran solution and an 8wt% N-methylimidazole-tetrahydrofuran solution, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants; 5) Scrape an oxidant solution with an iodine content of 6wt%, pyridine of 12wt%, tetrahydrofuran of 50wt% and the remainder of water, and rinse with a large amount of acetonitrile after sufficient reaction to remove the unreacted reactants. The characteristic wavelength of the auxiliary light in the above steps is 1480-1500nm.

[0077] Example 4

[0078] Substrate and surface pyramidal microstructure:

[0079] The substrate is a glass substrate, and the surface consists of a pyramidal recessed structure array with an upper side length of 500 μm, a recessed edge thickness of 15 μm, a bottom side length of 100 μm, and a height of 30 μm. The angle of the pyramid is 45 degrees, and there are micropores at the bottom of the recess with a pore diameter of 30 μm. The substrate is ultrasonically cleaned with ethanol-acetone-isopropanol in sequence.

[0080] Preparation of inverse opal biomimetic nanostructures:

[0081] A solution consisting of 0.3wt% styrene-acrylic acid-methyl methacrylate microspheres, 0.2wt% amorphous carbon and the remainder of water was coated on the surface of the cleaned substrate and cured at 150°C for 10 min; the microspheres were dissolved in toluene and then sintered at 200°C for 15 min; the diameter of the prepared inverse opal bionic nanostructure was 750-800nm.

[0082] Synthetic DNA:

[0083] 1) Scrape a universal solid phase carrier on the surface of the substrate and sinter at 180°C for 6 minutes; 2) After the substrate is cooled to room temperature, 8wt% trichloroacetic acid-dichloromethane solution is scraped, and after sufficient reaction, it is rinsed with a large amount of acetonitrile to remove the unreacted reactants; 3) 15wt% acetonitrile solution of ATCG base monomer and 10wt% 5-ethylmercaptotetrazole-acetonitrile solution are alternately scraped in sequence, and after sufficient reaction, it is rinsed with a large amount of acetonitrile to remove the unreacted reactants; 4) 20wt% acetic anhydride-tetrahydrofuran solution and 6wt% N-methylimidazole-tetrahydrofuran solution are scraped, and after sufficient reaction, it is rinsed with a large amount of acetonitrile to remove the unreacted reactants; 5) Scrape an oxidant solution with an iodine content of 8wt%, pyridine of 10wt%, tetrahydrofuran of 55wt% and the remainder of water, and after sufficient reaction, it is rinsed with a large amount of acetonitrile to remove the unreacted reactants. The characteristic wavelength of the auxiliary light in the above steps is 1480-1500nm.

[0084] Example 5

[0085] Substrate and surface pyramidal microstructure:

[0086] The substrate is a quartz substrate, and the surface consists of a pyramidal recessed structure array with an upper side length of 800 μm, a bottom side length of 150 μm, a recessed edge thickness of 8 μm, and a height of 30 μm. The angle of the pyramid is 30 degrees, and there are micropores at the bottom of the recess with a pore diameter of 50 μm. The substrate is ultrasonically cleaned in sequence with ethanol-acetone-isopropanol.

[0087] Preparation of inverse opal biomimetic nanostructures:

[0088] A solution consisting of 5wt% titanium dioxide microspheres, 5wt% polyurethane acrylic resin and the remainder of ethylene glycol methyl ether was coated on the surface of the cleaned substrate and cured at 120°C for 10 min; the microspheres were dissolved in toluene and then sintered at 180°C for 15 min; the diameter of the prepared inverse opal bionic nanostructure was 550-600nm.

[0089] Synthetic DNA:

[0090] 1) Apply a universal solid phase carrier on the surface of the substrate and sinter at 160°C for 7 minutes; 2) After the substrate is cooled to room temperature, apply a 9wt% trichloroacetic acid-dichloromethane solution, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants; 3) Apply a 15wt% acetonitrile solution of ATCG base monomer and an 8wt% 5-ethylmercaptotetrazole-acetonitrile solution alternately in sequence, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants; 4) Apply a 20wt% acetic anhydride-tetrahydrofuran solution and a 5wt% N-methylimidazole-tetrahydrofuran solution, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants; 5) Apply an oxidant solution with an iodine content of 8wt%, pyridine of 12wt%, tetrahydrofuran of 60wt% and the remainder of water, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants. The characteristic wavelength of light in the above steps of the reaction is 400-600nm.

[0091] Comparative Example 1

[0092] Substrate and surface pyramidal microstructure:

[0093] The substrate is a silicon substrate, and the surface is composed of a pyramidal recessed structure array with an upper side length of 100 μm, a recessed edge thickness of 10 μm, a bottom side length of 20 μm, and a height of 10 μm. The angle of the pyramid is 60 degrees, and there are micropores at the bottom of the recess with a pore diameter of 10 μm. The substrate is ultrasonically cleaned in sequence with ethanol-acetone-isopropanol.

[0094] Synthetic DNA:

[0095] 1) Apply a universal solid phase carrier on the surface of the substrate and sinter at 150°C for 8 minutes; 2) After the substrate is cooled to room temperature, apply a 10wt% trichloroacetic acid-dichloromethane solution, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants; 3) Apply a 10wt% acetonitrile solution of ATCG base monomer and a 10wt% 5-ethylmercaptotetrazole-acetonitrile solution alternately in sequence, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants; 4) Apply a 15wt% acetic anhydride-tetrahydrofuran solution and an 8wt% N-methylimidazole-tetrahydrofuran solution, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants; 5) Apply an oxidant solution with an iodine content of 6wt%, pyridine of 12wt%, tetrahydrofuran of 50wt% and the remainder of water, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants.

[0096] The above examples and comparative examples only illustrate the preparation process, and the cutting and purification of DNA use general methods, which are not limited by the present invention. Compared with the DNA prepared in Examples 1 to 5 of the present invention and Comparative Example 1 (see Table 1), the throughput is significantly improved.

[0097] Table 1 Comparison of DNA synthesis flux results between the embodiment and the comparative example (same reaction area)

[0098] Example Flux 1 2 million level 2 1 million level 3 1.2 million level 4 1.5 million level 5 1 million level Comparative Example 1 100,000 level

[0099] The above description is only a preferred embodiment of the present invention, and does not limit the patent scope of the present invention. All equivalent changes made using the present invention are within the patent protection scope of the present invention.

Claims

1. An application of a bionic nano-microstructure in DNA synthesis, characterized in that: S1 The bionic nano-microstructure is prepared by the following bionic nano-microstructure preparation method: (1) Substrate and surface pyramidal microstructure An array of pyramidal depressions is constructed on the surface of the substrate by photolithography; (2) Preparation of inverse opal biomimetic nanostructures preparing biomimetic nano-microstructures in the depressions; S2 DNA synthesis, including the following steps: 1) Fill the inverse opal structure with a universal solid phase carrier by scraping, and sinter at 120-200°C after filling, with a sintering time of 5-10 minutes; 2) After cooling to room temperature, the microwells are sequentially filled with deprotecting agent, ATCG base monomer, activator, capping agent, and oxidant to synthesize DNA. The characteristic wavelength of the auxiliary light for each reaction is 350-2000nm, and washing is required after the reaction.

2. The use of the bionic nano-microstructure in DNA synthesis according to claim 1, characterized in that: The deprotecting agent is a trichloroacetic acid-dichloromethane solution, the solution composition is 5-15wt% trichloroacetic acid and the balance dichloromethane; The activator is a 5-ethylmercaptotetrazole-acetonitrile solution, the solution composition is 5-15wt% 5-ethylmercaptotetrazole and the balance acetonitrile; The capping agent is acetic anhydride-tetrahydrofuran solution and N-methylimidazole-tetrahydrofuran solution, and the solution composition is 10-25wt% of acetic anhydride, 5-10wt% of N-methylimidazole and the balance of tetrahydrofuran; The oxidant is a solution of iodine-pyridine-tetrahydrofuran-water, wherein the oxidant solution contains 5-10wt% iodine, 10-15wt% pyridine, 40-60wt% tetrahydrofuran and the balance is water.

3. The use of the bionic nano-microstructure in DNA synthesis according to claim 1, characterized in that: The specific reaction steps of the DNA synthesis are: 1) Filling the inverse opal structure with a universal solid phase carrier by scraping, and sintering after filling; 2) After the substrate is cooled to room temperature, a deprotection agent solution is applied by scraping, and after sufficient reaction, it is rinsed with a large amount of acetonitrile to remove the unreacted reactants; 3) sequentially scraping an acetonitrile solution containing 1-20 wt% ATCG base monomer and an activator solution, and after sufficient reaction, washing with a large amount of acetonitrile to remove unreacted reactants; 4) Scrape the capping agent solution, rinse with a large amount of acetonitrile after sufficient reaction, and remove the unreacted reactants; 5) Scrape the oxidant solution, rinse with a large amount of acetonitrile after sufficient reaction to remove the unreacted reactants.

4. The use of the bionic nano-microstructure in DNA synthesis according to claim 1, characterized in that: The bionic nanostructure is an inverse opal structure, and its preparation process is: firstly, a coating that can form an opal structure is scraped on the surface of the cleaned substrate, and then the coating is solidified, the opal structure is dissolved, and sintered to form an inverse opal nanostructure with higher porosity.

5. The use of the bionic nano-microstructure in DNA synthesis according to claim 4, characterized in that: The array on the surface of the substrate is composed of pyramidal depressions with an upper side length of 5-1000 μm, an edge depression thickness of 1-20 μm, a bottom side length of 1-500 μm, and a height of 0.05-50 μm. The angle of the pyramid ranges from 0.5 to 90 degrees, and there are micropores at the bottom of the depression with a pore size of 1 to 100 μm. The substrate is any one of a silicon substrate, a silicon dioxide substrate, a quartz substrate or a glass substrate; The cleaning is to ultrasonically clean the substrate in sequence using ethanol, acetone, and isopropanol.

6. The use of the bionic nano-microstructure in DNA synthesis according to claim 4, characterized in that: The coating is composed of organic microspheres or inorganic microspheres, a supporting material and a solvent; The organic microspheres are composed of any one of polystyrene, styrene-acrylic acid copolymer, styrene-methyl acrylate copolymer, styrene-butyl acrylate copolymer, styrene-methacrylic acid copolymer, styrene-methyl methacrylate copolymer, styrene-butyl methacrylate copolymer, styrene-acrylic acid-methyl acrylate copolymer, styrene-acrylic acid-methyl methacrylate copolymer, styrene-acrylic acid-butyl acrylate copolymer, and styrene-acrylic acid-butyl methacrylate copolymer; The inorganic microsphere composition is selected from any one of silicon dioxide and titanium dioxide.

7. The use of the bionic nano-microstructure in DNA synthesis according to claim 6, characterized in that: The mass composition ratio of the organic or inorganic microspheres to the supporting material in the coating is 1:5-5:1; The supporting material is selected from any one or more of amorphous carbon, phenolic resin, epoxy resin, polystyrene, polyurethane, epoxy-acrylic resin, polyurethane acrylic resin, and phenolic epoxy acrylic resin; The coating solvent is selected from any one or more of water, ethanol, ethyl acetate, 6-caprolactone, butyl acetate, ethylene glycol methyl ether, ethylene glycol butyl ether, ethylene glycol phenyl ether, and propylene glycol methyl ether acetate.

8. The use of the bionic nano-microstructure in DNA synthesis according to claim 4, characterized in that: The curing is thermal curing, the heating temperature is 80-150°C, and the curing time is 5-15 minutes; the dissolution is to dissolve the organic or inorganic microspheres with toluene or concentrated sulfuric acid; The sintering temperature is 100-250°C and the sintering time is 10-20 minutes.

9. The use of the bionic nano-microstructure in DNA synthesis according to claim 4, characterized in that: The diameter of the inverse opal structure ranges from 500 to 1000 nm.

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