Preparation method and application of souvenir based on living body tissue DNA vitrification

Abstract

The invention belongs to the technical field of funeral services, and provides a souvenir preparation method and application based on life body tissue DNA vitrification, and the method comprises the following steps: extracting DNA from a life body tissue sample, and purifying to obtain a high-purity DNA solution; the preparation method comprises the following steps: mixing tetraethoxysilane, a catalyst, a solvent and water, and hydrolyzing to obtain silicon dioxide sol; the preparation method comprises the following steps: mixing a high-purity DNA (deoxyribonucleic acid) solution, silicon dioxide sol and 3-aminopropyltriethoxysilane to obtain DNA-silicon dioxide gel; and drying the DNA-silicon dioxide gel, combining the dried DNA-silicon dioxide gel with a base material, and preparing a commemorative product through a 3D printing, mold forming or engraving process. DNA molecules are embedded into a silicon dioxide gel matrix through electrostatic interaction and hydrogen bonds to form a transparent nano-composite, the transparent nano-composite is dried to obtain the DNA quartz glass, DNA information of the departed can be permanently stored, and the DNA quartz glass has extremely high memorial significance and emotional value.

Description

Technical Field

[0001] The present invention relates to the technical field of funeral services, and in particular to a method for preparing a souvenir based on the vitrification of DNA in living tissue and its application. Background Art

[0002] Traditional funeral services offer a limited range of commemorative options, making them difficult to meet people's growing demand for personalized and emotional remembrance. While recent technological advancements have led to the creation of memorabilia such as diamonds and jewelry, these technologies are costly and primarily serve decorative and emotional purposes, failing to preserve a person's DNA.

[0003] DNA, as the genetic material of life, carries unique information about an individual. Combining DNA with funeral services can create funeral mementos with greater commemorative significance and sentimental value. Therefore, a method for preparing and applying mementos based on the vitrification of DNA from living tissue is urgently needed. Summary of the Invention

[0004] The purpose of the present invention is to provide a souvenir preparation method and application based on the vitrification of DNA in living tissues, so as to overcome the defects in the prior art.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The present invention provides a method for preparing a souvenir based on the vitrification of DNA from living tissue, comprising the following steps:

[0007] S1. Extract DNA from living tissue samples and purify them to obtain a high-purity DNA solution;

[0008] S2, mixing tetraethoxysilane, a catalyst, a solvent and water and hydrolyzing them to obtain a silica sol;

[0009] S3, mixing a high-purity DNA solution, silica sol, and 3-aminopropyltriethoxysilane to encapsulate the DNA molecules in the silica sol to obtain a DNA-silica gel;

[0010] S4. After the DNA-silica gel is dried, it is combined with a substrate and prepared into a commemorative product through 3D printing, mold forming or carving technology. After forming, the surface of the product is polished or glazed to enhance durability.

[0011] Preferably, the DNA extraction and purification in step S1 includes the following steps:

[0012] S1.1. Digest the tissue sample with proteinase K and lysis buffer and centrifuge to remove impurities.

[0013] S1.2. Purify the DNA by silica gel membrane adsorption or phenol-chloroform extraction, and finally dissolve it in TE buffer and adjust the concentration to 0.5-5 mg / mL. The pH value of the TE buffer is 7.0-8.0.

[0014] Preferably, in step S2, the molar ratio of tetraethoxysilane, catalyst, solvent and water is 1:0.1:3.5-4.5:2.5-3.5;

[0015] The catalyst in step S2 is one or both of hydrochloric acid and ammonia water, and the concentration of the catalyst is 0.05-0.15 mol / L;

[0016] The solvent in step S2 is anhydrous ethanol.

[0017] Preferably, the hydrolysis temperature in step S2 is 20-25° C., the hydrolysis time is 12-24 h, and the stirring rate of the hydrolysis is 300-500 rpm.

[0018] Preferably, the pH of the silica sol system in step S2 is 4-5.

[0019] Preferably, the amount of 3-aminopropyltriethoxysilane added in step S3 is 5 to 10% of the volume of tetraethoxysilane;

[0020] The mixing temperature in step S3 is 15-25° C., and the mixing time is 0.5-1.5 h.

[0021] Preferably, the drying temperature in step S4 is 20-40° C., and the drying time is 6-24 hours.

[0022] Preferably, the substrate in step S4 is one or more of porous glass, transparent resin, glass bottle and card shell, and the substrate needs to be pretreated before combining the DNA-silica gel and the substrate, including plasma cleaning or chemical activation treatment, to enhance the adhesion strength of the DNA-silica gel.

[0023] Preferably, the product in step S4 further comprises at least one of the following functional coatings:

[0024] an anti-ultraviolet coating to prevent DNA degradation;

[0025] A hydrophobic coating for enhancing environmental resistance;

[0026] A fluorescent marker layer is used to display DNA distribution under a specific light source.

[0027] Preferably, the souvenir is subjected to quality inspection after being formed, and the inspection includes detecting DNA integrity by ultraviolet spectrophotometer. The inspection may also include observing whether the interface between DNA-silica gel and substrate is free of cracks by scanning electron microscopy.

[0028] The present invention has the following beneficial effects:

[0029] 1. This invention provides a method for preparing and applying a souvenir based on the vitrification of DNA from living tissue. DNA molecules are embedded in a silica gel matrix through electrostatic interactions and hydrogen bonds to form a transparent nanocomposite. After drying, the resulting DNA quartz glass can permanently preserve the DNA information of the deceased, possessing extremely high commemorative significance and sentimental value.

[0030] 2. The present invention provides a method for preparing souvenirs based on the vitrification of DNA from living tissues and their application, which can be customized into souvenirs of various shapes and materials to meet the personalized needs of different groups of people;

[0031] 3. The present invention provides a method for preparing a souvenir based on the vitrification of DNA from living tissue and its application. The present invention is low-cost and easy to promote, allowing more people to enjoy the convenience brought by technology. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The present invention is a flow chart of the method for preparing a souvenir based on the vitrification of DNA from living tissue. DETAILED DESCRIPTION

[0033] Exemplary embodiments will be described in detail herein, examples of which are illustrated in the accompanying drawings. In the following description, when referring to the drawings, identical numerals in different figures represent identical or similar elements unless otherwise indicated. The embodiments described in the following exemplary embodiments are not intended to represent all embodiments consistent with the present disclosure. Rather, they are merely examples of devices consistent with certain aspects of the present disclosure, as detailed in the appended claims.

[0034] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments; based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0035] like Figure 1 As shown, this embodiment provides a method for preparing a souvenir based on vitrification of DNA from living tissue, comprising the following steps:

[0036] S1. Extract DNA from living tissue samples and purify them to obtain a high-purity DNA solution;

[0037] S2, mixing tetraethoxysilane, a catalyst, a solvent and water and hydrolyzing them to obtain a silica sol;

[0038] S3, mixing a high-purity DNA solution, silica gel, and 3-aminopropyltriethoxysilane to encapsulate the DNA molecules in the silica sol to obtain a DNA-silica gel;

[0039] S4. After the DNA-silica gel is dried, it is combined with a substrate and prepared into a commemorative product through 3D printing, mold forming or carving technology. After forming, the surface of the product is polished or glazed to enhance durability.

[0040] In the present invention, the DNA extraction and purification in step S1 includes the following steps:

[0041] S1.1. Digest the tissue sample with proteinase K and lysis buffer and centrifuge to remove impurities.

[0042] S1.2. Purify the DNA by silica gel membrane adsorption or phenol-chloroform extraction, and finally dissolve it in TE buffer. The concentration is preferably adjusted to 0.5-5 mg / mL, and the pH value of the TE buffer is preferably 7.0-8.0.

[0043] In the present invention, the molar ratio of tetraethoxysilane, catalyst, solvent and water in step S2 is 1:0.1:3.5-4.5:2.5-3.5, more preferably 1:0.1:3.6-4.4:2.6-3.4, and more preferably 1:0.1:3.8-4.2:2.8-3.2.

[0044] In the present invention, the catalyst in step S2 is preferably one or both of hydrochloric acid and ammonia water, and the concentration of the catalyst is preferably 0.05-0.15 mol / L, more preferably 0.07-0.13 mol / L, and more preferably 0.09-0.11 mol / L.

[0045] In the present invention, the solvent in step S2 is preferably anhydrous ethanol.

[0046] In the present invention, the hydrolysis temperature in step S2 is preferably 20-25°C, more preferably 21-24°C, and even more preferably 22-23°C.

[0047] In the present invention, the hydrolysis time is preferably 12 to 24 hours, more preferably 14 to 22 hours, and more preferably 16 to 20 hours.

[0048] In the present invention, the stirring rate of the hydrolysis is preferably 300 to 500 rpm, more preferably 320 to 480 rpm, and even more preferably 350 to 450 rpm.

[0049] In the present invention, the pH of the silica sol system in step S2 is preferably 4-5.

[0050] In the present invention, a small amount of 3-aminopropyltriethoxysilane (APTES) is added to the silica sol. APTES can enhance the electrostatic effect. At this time, the positively charged amino groups on the SiO2 surface can form a stronger electrostatic effect with the negatively charged phosphate backbone of DNA, thereby achieving DNA fixation.

[0051] In the present invention, the amount of 3-aminopropyltriethoxysilane added in step S3 is preferably 5-10% by volume of tetraethoxysilane, more preferably 6-9%, and even more preferably 7-8%.

[0052] In the present invention, the mixing temperature in step S3 is preferably 15 to 25°C, more preferably 16 to 24°C, and even more preferably 17 to 23°C.

[0053] In the present invention, the mixing time is preferably 0.5 to 1.5 h, more preferably 0.6 to 1.4 h, and even more preferably 0.7 to 1.3 h.

[0054] In the present invention, the drying temperature in step S4 is preferably 20-40°C, more preferably 22-38°C, and even more preferably 25-35°C.

[0055] In the present invention, the drying time is preferably 6 to 24 hours, more preferably 8 to 22 hours, and even more preferably 10 to 20 hours.

[0056] In the present invention, ultra-long-term preservation is achieved by embedding DNA into a silica matrix, which has the characteristics of resistance to high temperature, oxidation and degradation. The dried DAN-silica gel has high transparency and a hardness close to that of glass.

[0057] In the present invention, the substrate in step S4 is one or more of porous glass, transparent resin, glass bottle and card shell. Before combining the DNA-silica gel and the substrate, the substrate needs to be pretreated, including plasma cleaning or chemical activation treatment, to enhance the adhesion strength of the DNA-silica gel.

[0058] In the present invention, the product in step S4 further comprises at least one of the following functional coatings:

[0059] an anti-ultraviolet coating to prevent DNA degradation;

[0060] A hydrophobic coating for enhancing environmental resistance;

[0061] A fluorescent marker layer is used to display DNA distribution under a specific light source.

[0062] In the present invention, the souvenir needs to be quality tested after being formed. The testing content includes testing the integrity of the DNA by ultraviolet spectrophotometer. The bonding interface between the DNA-silica gel and the substrate can also be observed by scanning electron microscopy to see whether there are any cracks.

[0063] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0064] Example 1

[0065] like Figure 1 As shown, the present invention's method for preparing a souvenir based on DNA mineralization technology for living tissue involves collecting approximately 0.1g of a hair sample from the deceased, placing it in a centrifuge tube, adding 500μL of lysis buffer (containing 1% SDS, 100mM NaCl, 10mM Tris-HCl, and 1mM EDTA, pH 8.0) and 20μg / mL proteinase K, and digesting the sample in a 50°C water bath for 2 hours, with vortexing every 30 minutes. After digestion, the sample is centrifuged at 12,000rpm for 10 minutes, the supernatant is transferred to a new centrifuge tube, and DNA is purified using silica gel membrane adsorption. The supernatant is transferred to a silica gel membrane centrifuge column, centrifuged at 8,000rpm for 1 minute, and the filtrate is discarded. The sample is then washed twice with 700μL of wash buffer (containing 80% ethanol, 10mM Tris-HCl, and 1mM EDTA, pH 8.0), centrifuged at 8,000rpm for 1 minute each time, and the filtrate is discarded. Finally, place the spin column in a new centrifuge tube and add 50 μL of TE buffer (pH 8.0). Let it stand at room temperature for 1 minute, then centrifuge at 12,000 rpm for 1 minute. Collect the eluate to obtain the purified DNA solution. Measure the absorbance of the DNA solution at 260 nm and 280 nm using a UV spectrophotometer. The calculated A260 / A280 ratio is 1.8, indicating good DNA purity. Adjust the DNA solution concentration to 1 mg / mL.

[0066] Tetraethoxysilane, 0.1M hydrochloric acid, anhydrous ethanol and water were mixed in a molar ratio of 1:0.1:4:3, and hydrolysis reaction was carried out at 20°C for 12 hours. HCl was used to maintain the pH of the hydrolyzate at 4-5 to obtain silica sol. DAN solution and APTES (the added amount was 10% of the volume of tetraethoxysilane) were slowly added to the silica gel, and stirred at 20°C at a rate of 100 rpm for 1 hour to allow the DNA molecules to be wrapped in the silica sol to obtain DNA-silica gel. The DNA-silica gel was dried and solidified, and dried at 30°C for 10 hours to obtain a solid transparent substance with a hardness close to that of glass.

[0067] Mix the dried DNA-silica gel with a transparent resin in a mass ratio of 1:10, stir thoroughly, and pour into the hopper of a light-curing 3D printer. Import the 3D pendant model file, set the layer thickness to 0.05 mm, and the print speed to 20 mm / s, and start printing. After printing, remove the pendant blank and clean it with isopropyl alcohol to remove any uncured resin. Then, polish the pendant blank with 400#, 800#, and 1200# sandpaper, followed by polishing with 1.5μm diamond polishing paste to a mirror finish.

[0068] A laser engraving machine was used to engrave the deceased's name "Zhang San" and date of birth and death "1950.05.21-2025.05.21" on the surface of the pendant. The engraving power was 5W and the engraving speed was 100mm / s. The deceased's photo was printed on the back of the pendant using UV printing technology with a printing resolution of 600dpi.

[0069] The pendant was immersed in an ethanol solution containing 2% titanium dioxide nanoparticles for 10 minutes, taken out, and dried at 60°C to form an anti-ultraviolet coating.

[0070] Weigh 5 g of glycerol, 0.372 g of EDTA·2Na, and 1.211 g of Tris and dissolve them in 100 mL of deionized water. Adjust the pH to 8.0 with hydrochloric acid and dilute to 1 L to prepare the preservation solution. Immerse the active DNA sample in the DNA-silica gel in the preservation solution, seal the container with nitrogen, and store in a refrigerator at 4°C.

[0071] DNA was extracted from the preservation solution and the human mitochondrial DNA control region fragment (about 500bp) was amplified by PCR. Gel electrophoresis showed a single bright band, indicating that the DNA integrity was good. The surface hardness of the pendant was measured to be 5H using a hardness tester. After the wear resistance test (RCA paper tape, 175g load, 100 cycles), there were no obvious scratches on the surface. After the corrosion resistance test (5% NaCl solution, 24 hours), there was no corrosion on the surface.

[0072] The pendant souvenir prepared in this embodiment not only carries the DNA information of the deceased and has unique commemorative significance, but can also serve as a carrier for a biological sample library.

[0073] Example 2

[0074] Approximately 0.1 g of hair sample was collected from the deceased and placed in a centrifuge tube. 500 μL of lysis buffer (containing 1% SDS, 100 mM NaCl, 10 mM Tris-HCl, and 1 mM EDTA, pH 8.0) and 20 μg / mL proteinase K were added. The sample was digested in a 50°C water bath for 2 hours, with vortex mixing every 30 minutes. After digestion, the sample was centrifuged at 12,000 rpm for 10 minutes, and the supernatant was transferred to a new centrifuge tube. DNA was purified using silica gel membrane adsorption. The supernatant was transferred to a silica gel membrane centrifuge column, centrifuged at 8,000 rpm for 1 minute, and the filtrate was discarded. The sample was washed twice with 700 μL of wash buffer (containing 80% ethanol, 10 mM Tris-HCl, and 1 mM EDTA, pH 8.0), centrifuged at 8,000 rpm for 1 minute each time, and the filtrate was discarded. Finally, place the spin column in a new centrifuge tube and add 50 μL of TE buffer (pH 8.0). Let it stand at room temperature for 1 minute, then centrifuge at 12,000 rpm for 1 minute. Collect the eluate to obtain the purified DNA solution. Measure the absorbance of the DNA solution at 260 nm and 280 nm using a UV spectrophotometer. The calculated A260 / A280 ratio is 1.8, indicating good DNA purity. Adjust the DNA solution concentration to 0.5 mg / mL.

[0075] Tetraethoxysilane, 0.1M hydrochloric acid, anhydrous ethanol and water were mixed in a molar ratio of 1:0.1:3.5:2.5, and the mixture was hydrolyzed at 20°C for 12 hours. HCl was used to maintain the pH of the hydrolyzate at 4-5 to obtain a silica sol. DAN solution and APTES (the added amount was 5% of the volume of tetraethoxysilane) were slowly added to the silica gel and stirred at 100 rpm at 15°C for 0.5 hours to allow the DNA molecules to be wrapped in the silica sol to obtain a DNA-silica gel. The DAN-silica gel was cast into a rectangular card slot mold, and then the DNA-silica gel was dried and solidified. It was dried at 20°C for 8 hours to obtain a solid transparent substance with a hardness close to that of glass.

[0076] A glass or plastic card case is used as the substrate and immersed in a plasma cleaner at 100W for 5 minutes in an oxygen atmosphere to remove surface impurities and enhance hydrophilicity. The dried, formed DNA-silica gel is placed inside the card case. Using an engraving machine, a personal message is engraved on the surface of the card case, completing the souvenir.

[0077] Example 3

[0078] Approximately 0.1 g of hair sample was collected from the deceased and placed in a centrifuge tube. 500 μL of lysis buffer (containing 1% SDS, 100 mM NaCl, 10 mM Tris-HCl, and 1 mM EDTA, pH 8.0) and 20 μg / mL proteinase K were added. The sample was digested in a 50°C water bath for 2 hours, with vortex mixing every 30 minutes. After digestion, the sample was centrifuged at 12,000 rpm for 10 minutes, and the supernatant was transferred to a new centrifuge tube. DNA was purified using silica gel membrane adsorption. The supernatant was transferred to a silica gel membrane centrifuge column, centrifuged at 8,000 rpm for 1 minute, and the filtrate was discarded. The sample was washed twice with 700 μL of wash buffer (containing 80% ethanol, 10 mM Tris-HCl, and 1 mM EDTA, pH 8.0), centrifuged at 8,000 rpm for 1 minute each time, and the filtrate was discarded. Finally, place the spin column in a new centrifuge tube and add 50 μL of TE buffer (pH 8.0). Let it stand at room temperature for 1 minute, then centrifuge at 12,000 rpm for 1 minute. Collect the eluate to obtain the purified DNA solution. Measure the absorbance of the DNA solution at 260 nm and 280 nm using a UV spectrophotometer. The calculated A260 / A280 ratio is 1.8, indicating good DNA purity. Adjust the DNA solution concentration to 0.5 mg / mL.

[0079] Tetraethoxysilane, 0.1M hydrochloric acid, anhydrous ethanol and water were mixed in a molar ratio of 1:0.1:4.5:3.5, and hydrolysis reaction was carried out at 25°C for 24 hours. HCl was used to maintain the pH of the hydrolyzate at 4-5 to obtain silica sol. DAN solution and APTES (the added amount was 10% of the volume of tetraethoxysilane) were slowly added to the silica gel and stirred at 200 rpm at 25°C for 1.5 hours to allow the DNA molecules to be wrapped in the silica sol to obtain DNA-silica gel. The DNA-silica gel was then dried and solidified, and dried at 20°C for 8 hours to obtain a solid transparent substance with a hardness close to that of glass.

[0080] A small glass bottle was selected as the substrate and immersed in a plasma cleaner under an oxygen atmosphere at 100W for 5 minutes to remove surface impurities and enhance hydrophilicity. The dried DNA was then placed in the small glass bottle. A personal engraving machine was then used to engrave the surface of the glass bottle, completing the souvenir.

[0081] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A method for preparing a souvenir based on the vitrification of DNA from living tissue, characterized in that: It includes the following steps: S1. Extract DNA from living tissue samples and purify them to obtain a high-purity DNA solution; S2, mixing tetraethoxysilane, a catalyst, a solvent and water and hydrolyzing them to obtain a silica sol; S3, mixing a high-purity DNA solution, silica sol, and 3-aminopropyltriethoxysilane to encapsulate the DNA molecules in the silica sol to obtain a DNA-silica gel; S4. After the DNA-silica gel is dried, it is combined with a substrate and prepared into a commemorative product through 3D printing, mold forming or carving technology. After forming, the surface of the product is polished or glazed to enhance durability.

2. The method for preparing a souvenir based on vitrification of DNA from living tissue according to claim 1, characterized in that: The DNA extraction and purification in step S1 includes the following steps: S1.

1. Digest the tissue sample with proteinase K and lysis buffer and centrifuge to remove impurities. S1.

2. Purify the DNA by silica gel membrane adsorption or phenol-chloroform extraction, and finally dissolve it in TE buffer and adjust the concentration to 0.5-5 mg / mL. The pH value of the TE buffer is 7.0-8.

0.

3. The method for preparing a souvenir based on vitrification of DNA from living tissue according to claim 1, characterized in that: In step S2, the molar ratio of tetraethoxysilane, catalyst, solvent and water is 1:0.1:3.5-4.5:2.5-3.5; The catalyst in step S2 is one or both of hydrochloric acid and ammonia water, and the concentration of the catalyst is 0.05-0.15 mol / L; The solvent in step S2 is anhydrous ethanol.

4. The method for preparing a souvenir based on vitrification of DNA from living tissue according to claim 1, characterized in that: The hydrolysis temperature in step S2 is 20-25° C., the hydrolysis time is 12-24 h, and the stirring rate of the hydrolysis is 300-500 rpm.

5. The method for preparing a souvenir based on vitrification of DNA from living tissue according to claim 1, characterized in that: The pH of the silica sol system in step S2 is 4-5.

6. The method for preparing a souvenir based on vitrification of DNA from living tissue according to claim 1, characterized in that: In step S3, the amount of 3-aminopropyltriethoxysilane added is 5-10% of the volume of tetraethoxysilane; The mixing temperature in step S3 is 15-25° C., and the mixing time is 0.5-1.5 h.

7. The method for preparing a souvenir based on vitrification of DNA from living tissue according to claim 1, characterized in that: The drying temperature in step S4 is 20-40° C., and the drying time is 6-24 hours.

8. The method for preparing a souvenir based on vitrification of DNA from living tissue according to claim 1, characterized in that: In step S4, the substrate is one or more of porous glass, transparent resin, glass bottle, and card case. Before combining the DNA-silica gel and the substrate, the substrate needs to be pretreated, including plasma cleaning or chemical activation treatment, to enhance the adhesion strength of the DNA-silica gel.

9. The method for preparing a souvenir based on vitrification of DNA from living tissue according to claim 1, characterized in that: The product in step S4 further includes at least one of the following functional coatings: an anti-ultraviolet coating to prevent DNA degradation; A hydrophobic coating for enhancing environmental resistance; A fluorescent marker layer is used to display DNA distribution under a specific light source.

10. The method for preparing a souvenir based on vitrification of DNA from living tissue according to claim 1, characterized in that: After the souvenir is formed, it needs to undergo quality inspection. The inspection content includes detecting the integrity of the DNA by ultraviolet spectrophotometer. The inspection can also be carried out by scanning electron microscopy to observe whether the interface between the DNA-silica gel and the substrate is free of cracks.

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