A method for preparing a hydrogel nucleic acid separation device

Abstract

The application belongs to the field of engineering technology and provides a preparation method of a nucleic acid separation device. The preparation method of the application prepares a hydrogel nucleic acid adsorption device through a photo-initiated radical polymerization reaction, avoiding the cumbersome limitations of traditional polymerization reactions which need to be carried out under anaerobic conditions. The preparation method is simple and fast, and does not depend on professional and precise instruments. The raw materials are cheap and easy to obtain. The method can efficiently load silica particles, effectively combine the plasticity of the hydrogel with the high specific surface area of the silica particles and the strong binding capacity of the nucleic acid, and further obtain a handheld nucleic acid extraction device which can change shape, size and quantity at any time according to application requirements, has a high specific surface area and excellent nucleic acid extraction performance. Compared with traditional processes, the preparation process is simpler, the application range is wider, and the application requirements of low resource environment are greatly met, and the application prospect in the field of outdoor and clinical nucleic acid extraction technology is broad.

Description

Technical Field

[0001] This invention belongs to the field of engineering technology and relates to a method for preparing a hydrogel nucleic acid separation device. Background Technology

[0002] Nucleic acids are macromolecules widely found in all living organisms, including two main categories: DNA and RNA. They play a decisive role in a series of major life phenomena such as growth, heredity, and variation. Methods for extracting nucleic acids include liquid-phase separation and solid-phase methods (magnetic separation and non-magnetic separation). Solid-phase separation generally uses materials with nucleic acid adsorption properties, such as magnetic beads, silica, cellulose, silica gel, or synthetic polymers specifically designed for nucleic acid extraction. Currently, the most commonly used methods are magnetic bead extraction (magnetic separation) and centrifugal column extraction (non-magnetic separation). The magnetic bead method used in application CN201710113885.X, entitled "A Method for Nucleic Acid Extraction by Magnetic Beads," separates nucleic acids by using silica-coated magnetic particles (positively charged) to adsorb nucleic acids (negatively charged). After repeated washing and agitation in a lysis buffer, a magnetic field is applied, and the magnetic beads adhere to the tube wall. The liquid is then removed, and the adsorption-washing-desorption steps are repeated. Application 202210295109.7 discloses a nucleic acid extraction centrifuge column, a nucleic acid extraction device, and a nucleic acid extraction method. This method uses a centrifuge column, often made of silicon-based or polymer materials, to extract nucleic acids through electrostatic interactions, hydrogen bonding, and intermolecular forces. Centrifugation is then used to separate the nucleic acid lysis buffer, washing buffer, and elution buffer by passing them through the adsorbent material. The two methods mentioned above are commonly used for nucleic acid separation. However, the magnetic bead method for magnetic separation relies on a magnetic field, which causes magnetic particles to agglomerate and settle. The non-magnetic centrifuge column method requires multiple auxiliary operations, relies on centrifugation technology, has a long separation time, low throughput, and introduces PCR inhibitory components during the process.

[0003] To address the aforementioned issues, the applicant previously disclosed a non-magnetic nucleic acid solid-phase separation technology based on 3D-printed functional devices. This technology enables high-throughput, high-quality extraction of DNA from biological samples efficiently and without relying on centrifugation or external magnetic fields. However, the fabrication of 3D-printed devices depends on high-precision, expensive 3D printers, requires highly skilled personnel, and has a long development cycle for inks compatible with 3D printing, making it unsuitable for low-resource areas. Therefore, developing a simple, instrument-free method for fabricating nucleic acid extraction devices is particularly important for low-resource regions. Summary of the Invention

[0004] This invention provides a novel method for preparing nucleic acid separation devices. The preparation method is simple and rapid, and the device shape can be flexibly adjusted as needed to solve the problems of traditional nucleic acid separation relying on centrifugation and low throughput. This method is suitable for preparing nucleic acid separation devices when the laboratory does not have the equipment for material construction.

[0005] The technical solution of the present invention:

[0006] A method for preparing a hydrogel nucleic acid separation device, comprising the following steps:

[0007] (1) Add crosslinking agent and photoinitiator to hydrogel monomer to prepare prepolymer solution;

[0008] (2) After mixing the prepolymer liquid, add it to the mold, insert the handle, place it under the ultraviolet lamp for curing, and carry out free radical polymerization reaction under light conditions;

[0009] (3) Hold the handle to remove the device from the mold, wash and dry the device and then store it.

[0010] The hydrogel monomers are commonly used hydrogel monomers such as acrylic acid and acrylate; the crosslinking agents are commonly used crosslinking agents such as N'N-methylenebisacrylamide and polyethylene glycol diacrylate, and the crosslinking agents account for 2-50 wt% of the hydrogel monomers; the initiator is a photoinitiator such as (2,4,6-trimethylbenzoyl) diphenylphosphine oxide, and the amount added is 0.1-0.5 wt% of the hydrogel monomers.

[0011] Preferably, silica particles can be added to the prepolymer solution. The silica particles can be silica as a single component or other substances with silica as the main component, such as quartz or glass, and their size is not limited. Preferably, the silica particles are a mixture of 50-180 mesh quartz sand particles, and the amount of silica particles added is 20-50 wt% of the total solid-liquid mixture. At the same time, the particle structure can achieve a rough, frosted material on the surface of the hydrogel device.

[0012] Photoinitiated free radical polymerization is carried out at room temperature with a light wavelength of 280-400 nm and an irradiation time of 3-10 min. Inverting the mold ensures uniform cross-linking at the bottom of the device, and the irradiation time is 3-40 min. After demolding, the device is washed using one or more of water and ethanol as the washing solution, at least once, to remove excess monomers and impurities and reduce the risk of contamination in subsequent use. The washed device is then dried at 0-80°C for 3 min-24 h. The device is stored at room temperature and protected from light.

[0013] The aforementioned mold is a cavity-type container designed to meet the needs of subsequent DNA extraction; preferred options include centrifuge tubes, eight-tube arrays, 96-well plates, and deep-well plates. If a handheld device is to be fabricated, the handle is rod-shaped, with no restrictions on shape or material.

[0014] The beneficial effects of this invention are:

[0015] (1) This invention prepares hydrogel nucleic acid adsorption devices by photo-initiated free radical polymerization, avoiding the cumbersome and complicated polymerization under anaerobic conditions in traditional processes. Compared with centrifugal column-type and magnetic particle-type nucleic acid extraction carriers and devices, the raw materials are simpler, cheaper and more readily available, the preparation process is simple and fast, and it does not rely on other professional materials to construct equipment, making it suitable for low-resource areas and outdoor applications.

[0016] (2) The device uses hydrogel as the substrate. The surface groups of the hydrogel polymer material are rich, which can increase the electrostatic interaction, hydrogen bonding and intermolecular forces of nucleic acid adsorption to achieve efficient separation of nucleic acid. By loading silica particles, the plasticity of the hydrogel material and the high nucleic acid adsorption of silica are combined to further improve the nucleic acid adsorption performance.

[0017] (3) This preparation method is simple and easy to implement, does not rely on machines, and can efficiently load functional particles. It solves the problem that in traditional hydrogel molding processes such as 3D printers, the doping of solid particles in the printing ink can easily damage the machine. At the same time, the doped particles increase the surface roughness and specific surface area, further increasing the nucleic acid binding sites and improving the nucleic acid extraction performance of the device.

[0018] (4) This preparation method can be used to prepare nucleic acid extraction equipment of different quantities, sizes and shapes by changing the mold, and can be used with different containers, making it more widely applicable and flexible.

[0019] (5) The device prepared in this way has a simple extraction process for nucleic acid extraction and does not rely on instruments. Compared with traditional centrifugal column and magnetic separation nucleic acid extraction methods, it does not rely on professional equipment and does not have the risk of particle aggregation and sedimentation, which greatly meets the needs of low-resource environment, outdoor and clinical nucleic acid extraction applications. Attached Figure Description

[0020] Figure 1 It is a hydrogel nucleic acid adsorption device; (a) a single nucleic acid extraction device; (b) a nucleic acid extraction device adapted to an eight-tube array; (c) a top view of a nucleic acid extraction device adapted to a 1.5mL centrifuge tube; (d) a side view of a nucleic acid extraction device adapted to a 1.5mL centrifuge tube.

[0021] Figure 2 These are electrophoresis results for devices using DMAM as the crosslinking agent. 1-3: Three parallel results for devices without added quartz sand; 4-6: Three parallel results for devices with added quartz sand; M: DNA Maker; NC: Negative control using water as a template.

[0022] Figure 3The nucleic acid adsorption performance of the device with PEGDA as the crosslinking agent is shown in the figures. Among them, (a) the nucleic acid extraction concentration of the device with and without silica (PAA) is shown in the figures; (b) the A260 / A280 and A260 / A230 of the device with and without silica (PAA) is shown in the figures.

[0023] Figure 4 The nucleic acid adsorption performance of the nucleic acid extraction device matched with the eight-tube array is shown in the figure; (a) the nucleic acid extraction concentration of the eight devices; and (b) the nucleic acid extraction A260 / A280 and A260 / A230 of the eight devices.

[0024] Figure 5 The nucleic acid adsorption performance of the centrifuge tube-type nucleic acid extraction device includes (a) nucleic acid extraction concentrations with different silica addition amounts; and (b) A260 / A280 and A260 / A230 with different silica addition amounts.

[0025] Figure 6 The nucleic acid adsorption performance of devices with different silica addition amounts; (a) nucleic acid extraction concentration of devices with different silica addition amounts; (b) A260 / A280 and A260 / A230 of devices with different silica addition amounts. Detailed Implementation

[0026] The following are specific embodiments of the present invention, which are described in conjunction with the accompanying drawings to further illustrate the technical solutions of the present invention. However, the present invention is not limited to these embodiments.

[0027] The following description, in conjunction with the technical solutions, further illustrates the content of this invention. It should be noted that the following descriptions are exemplary and intended to provide further explanation of the invention. Unless otherwise stated, all scientific and technical terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0028] Unless otherwise specified, the materials, practices, and experimental equipment involved in the embodiments of this invention are all commercially available products in the relevant chemical and biotechnology fields.

[0029] Example 1

[0030] (1) Take 2 ml of acrylic monomer and 0.05 g of crosslinking agent N'N-methylenebisacrylamide (DMAM), and then add 0.002 g of photoinitiator and deionized water to prepare a prepolymer solution;

[0031] (2) The prepolymer solution in step (1) is divided into two groups. One group is mixed with 1g of silica particles of 50-80 mesh quartz sand and 120-180 mesh quartz sand, and stirred evenly. The other group is not mixed with quartz sand.

[0032] (3) After mixing, add (2) to a single 0.2 mL centrifuge tube, add 80 μL of solid solution mixture to each well, and mix well;

[0033] (4) Insert a toothpick as a handle into the centrifuge tube in step (3), place it under a 400nm ultraviolet lamp for 5 minutes to cure it, and carry out free radical polymerization under light conditions;

[0034] (5) Take out the centrifuge tube from (4), invert it and irradiate the bottom for 3 minutes, and irradiate the side for 3 minutes;

[0035] (6) Hold the handle to remove the device from the centrifuge tube and wash it three times with anhydrous ethanol.

[0036] (7) Dry at 50℃ for 10 minutes, then store away from light for later use (e.g.) Figure 1 a).

[0037] The obtained polyacrylic acid-doped silica nucleic acid extraction device was used to extract nucleic acids from biological samples. The process consisted of four steps: lysis, adsorption, washing, and elution.

[0038] (1) Prepare nucleic acid CTAB lysis buffer and mix by centrifugation.

[0039] (2) Take the biological sample to be tested and place it in a 1.5 mL centrifuge tube. Add 400 μL of lysis buffer and 20 μL of proteinase K and shake well.

[0040] (3) Place at 60℃ for 60 min until the biological sample tissue is completely dissolved and the solution becomes clear;

[0041] (4) Take 100 μL of the lysed solution and add 60 μL of isopropanol into a 0.2 mL centrifuge tube. Insert the device into the centrifuge tube and shake it up and down gently for 1 minute before taking it out.

[0042] (5) Take a new 0.2 mL centrifuge tube and add it to 150 μL of washing solution. Add the device that has been adsorbed with nucleic acid into the solution and shake it gently up and down for 30 seconds. Then take it out and let it air dry at room temperature.

[0043] (6) Take a new 0.2 mL centrifuge tube, add 80 μL of elution buffer and the dried device, and shake up and down for 1 min to elute the nucleic acid from the device.

[0044] The obtained nucleic acid samples were amplified by PCR and then subjected to electrophoresis. The results proved (e.g.) Figure 2This invention utilizes photo-initiated free radical polymerization to prepare hydrogel nucleic acid adsorption devices, avoiding the cumbersome and complex polymerization process under anaerobic conditions in traditional methods. The raw materials are simple, inexpensive, and readily available; the preparation process is simple and rapid, requiring no additional specialized materials for equipment construction; and the prepared devices can selectively separate nucleic acids from mixed biological samples, making them suitable for low-resource areas and outdoor applications.

[0045] This embodiment also illustrates that the preparation method can efficiently load functional particles without relying on machines, solving the problem of machine damage caused by doping solid particles into printing ink in traditional hydrogel molding processes such as 3D printers. Simultaneously, the increased surface roughness and specific surface area due to the doped particles further enlarge nucleic acid binding sites, improving the nucleic acid extraction performance of the device. Figure 3 ).

[0046] Example 2

[0047] (1) Take 2 mL of acrylic monomer and 1 mL of crosslinking agent polyethylene glycol diacrylate, and add 0.004 g of photoinitiator TPO to prepare a prepolymer solution;

[0048] (2) Add 1g of 50-80 mesh quartz sand and 1g of 120-180 mesh quartz sand to the prepolymer solution in step (1) and stir evenly;

[0049] (3) After mixing, add (2) to the eight-tube array, add 80 μL of solid liquid mixture to each hole, and mix well;

[0050] (4) In step (3), the eight-tube array is inserted into the comb-shaped handle and placed under a 300nm ultraviolet lamp for 5 minutes to cure and carry out free radical polymerization under light conditions.

[0051] (5) Take out the eight-tube array from (4), invert it and irradiate the bottom for 10 minutes, and irradiate the side for 10 minutes;

[0052] (6) Hold the handle to remove the device from the eight-tube array, and wash it three times with deionized water and anhydrous ethanol; (7) Dry at 25°C for 24 hours and then store it away from light for later use (e.g. Figure 1 b).

[0053] The obtained nucleic acid extraction device was used to extract nucleic acids from biological samples, and the biological samples were lysed using commercially available nucleic acid lysis buffer.

[0054] (1) Take the biological sample to be tested and place it in a 1.5 mL centrifuge tube. Add 400 μL of lysis buffer and 20 μL of proteinase K and shake well.

[0055] (2) Place at 56℃ for 40 minutes until the biological sample tissue is completely dissolved and the solution becomes clear;

[0056] (3) Take 100 μL of the lysed solution and add 60 μL of isopropanol into an eight-tube bundle;

[0057] (4) Insert the device into the above eight-tube array and gently shake it up and down for 1 minute before taking it out.

[0058] (5) Take a new 0.2 mL eight-tube and add 150 μL of 75 wt% ethanol. Add the device with the adsorbed nucleic acid into it and shake it gently up and down for 30 seconds. Then take it out and let it air dry at room temperature.

[0059] (6) Take a new eight-tube strip, add 80 μL of TE elution buffer and the dried device, and shake it up and down for 1 minute to elute the nucleic acid from the device.

[0060] The obtained nucleic acid samples were quantified using a UV spectrophotometer, and the extracted nucleic acid concentrations were 45.77 ng / μL, 37.96 ng / μL, 31.78 ng / μL, 26.78 ng / μL, 25.01 ng / μL, 30.59 ng / μL, 37.95 ng / μL, and 33.41 ng / μL, respectively. The results demonstrate that this preparation method can achieve the preparation of nucleic acid extraction devices of different quantities, sizes, and shapes by changing the mold, and can be used with different containers, making it more versatile and applicable to a wider range of scenarios. Devices prepared using 50 wt% polyethylene glycol diacrylate as a crosslinking agent and 40 wt% silica as an additive exhibit high nucleic acid extraction yields.

[0061] Example 3

[0062] (1) Take 2 mL of acrylic monomer and 0.1 g of crosslinking agent N'N methylenebisacrylamide (PAA-SiO2-1), and another group uses 0.2 g of crosslinking agent N'N methylenebisacrylamide (PAA-SiO2-2). Add 0.004 g of photoinitiator and mix with deionized water to prepare a prepolymer solution.

[0063] (2) Add 1g of 50-80 mesh quartz sand and 1g of 120-180 mesh quartz sand to the prepolymer solution in step (1) and stir evenly;

[0064] (3) After mixing, add (2) to a 1.5 mL centrifuge tube, add 200 μL of the above prepolymer to each well, and insert a new 1.5 mL centrifuge tube so that the solid-liquid mixture is "squeezed" onto the wall.

[0065] (4) The centrifuge tubes from step (3) were placed under a 300nm UV lamp for 5 minutes to cure and undergo free radical polymerization under light conditions.

[0066] (5) Take out the two centrifuge tubes from (4), invert them and irradiate the bottom for 3 minutes and the side for 3 minutes;

[0067] (6) Remove the upper centrifuge tube and wash the inner wall of the centrifuge tube three times with deionized water;

[0068] (7) Dry at 80℃ for 3 minutes, then store away from light for later use (e.g.) Figure 1 c, d).

[0069] The obtained nucleic acid extraction device was used to extract nucleic acids from biological samples, and the biological samples were lysed using commercially available nucleic acid lysis buffer.

[0070] (1) Take the biological sample to be tested and place it in a 1.5 mL centrifuge tube. Add 400 μL of lysis buffer and 20 μL of proteinase K and shake well.

[0071] (2) Place at 60℃ for 60 minutes until the sample tissue is completely dissolved and the solution becomes clear;

[0072] (3) Take 200 μL of the lysed solution and add 100 μL of anhydrous ethanol into a centrifuge tube, and gently shake the centrifuge tube for 1 min.

[0073] (4) Add 300 μL of 75 wt% ethanol to the centrifuge tube in step (3), shake gently up and down for 30 seconds, then remove and air dry at room temperature;

[0074] (5) Take 100 μL of TE elution buffer from the centrifuge tube in (4) and shake it up and down for 1 min to elute the nucleic acid from the device.

[0075] The obtained nucleic acid samples were tested using a UV spectrophotometer to determine their nucleic acid adsorption content, A260 / A280, and A260 / A230 ratios. The results demonstrate (e.g.) Figure 5 The centrifuge tube-type nucleic acid extraction device prepared by this method has the ability to separate nucleic acids, proving that this method of preparing nucleic acid extraction devices with acrylic acid-doped quartz sand has high flexibility and can be used to prepare corresponding nucleic acid extraction devices with different containers according to different experimental requirements.

[0076] Example 4

[0077] (1) Take 2 mL of acrylic monomer, 1 mL of crosslinking agent polyethylene glycol diacrylate and 1 g of N'N methylene bisacrylamide, add 0.001 g of photoinitiator, and prepare a prepolymer solution;

[0078] (2) Step (1) is divided into four groups. The first group of prepolymer liquid does not add quartz sand (PAA). The second group adds 2g of 120-180 mesh quartz sand (PAA-SiO2-small). The third group adds 2g of 50-80 mesh quartz sand (PAA-SiO2-large). The last group adds 1g of 50-80 mesh quartz sand and 1g of 120-180 mesh quartz sand (PAA-SiO2-mixed) and stirs evenly.

[0079] (3) After mixing, add (2) to the eight-tube array, add 80 μL of solid liquid mixture to each hole, and mix well;

[0080] (4) Insert the 3D-printed handle into the eight-tube array in step (3), place it under a 300nm ultraviolet lamp for curing for 40 minutes, and carry out free radical polymerization under light conditions;

[0081] (5) Take out the eight-tube array from (4), invert it and irradiate the bottom for 40 minutes, and irradiate the side for 40 minutes;

[0082] (6) Hold the handle to remove the device from the eight-tube strip, and wash it three times with deionized water and anhydrous ethanol; (7) Dry it at 25°C for 1 hour and then store it in the dark for later use.

[0083] The obtained nucleic acid extraction device was used to extract nucleic acids from a mixed biological sample, and the biological sample was lysed using a self-prepared lysis buffer.

[0084] (1) Take the biological sample to be tested and place it in a 1.5 mL centrifuge tube. Add 400 μL of lysis buffer and 20 μL of proteinase K and shake well.

[0085] (2) Place at 56℃ for 40 minutes until the biological sample tissue is completely dissolved and the solution becomes clear;

[0086] (3) Take 100 μL of the lysed solution and add 60 μL of isopropanol into an eight-tube bundle;

[0087] (4) Insert the device into the above eight-tube array and gently shake it up and down for 1 minute before taking it out.

[0088] (5) Take a new 0.2 mL eight-tube and add 150 μL of 75 wt% ethanol. Add the device with the adsorbed nucleic acid into it and shake it gently up and down for 30 seconds. Then take it out and let it air dry at room temperature.

[0089] (6) Take a new eight-tube strip, add 80 μL of TE elution buffer and the dried device, and shake it up and down for 1 minute to elute the nucleic acid from the device.

[0090] The obtained nucleic acid samples were tested using a UV spectrophotometer to determine their nucleic acid adsorption content, A260 / A280, and A260 / A230 ratios. The results demonstrate (e.g.) Figure 6 The four sets of nucleic acid extraction devices prepared by this method have the ability to separate nucleic acids. The nucleic acid extraction effect of the material doped with quartz sand is higher than that of the undoped material. The size of the doped particles is not limited and can be optimized and adjusted according to the experimental purpose.

[0091] Example 5

[0092] (1) Take 2 mL of acrylic monomer and 1 mL of crosslinking agent polyethylene glycol diacrylate, add 0.01 g of photoinitiator, and prepare a prepolymer solution;

[0093] (2) In step (1), add 1g of 50-80 mesh quartz sand and 1g of 120-180 mesh quartz sand to the prepolymer solution and stir until homogeneous;

[0094] (3) After mixing, add (2) to the eight-tube array, add 80 μL of solid liquid mixture to each hole, and mix well;

[0095] (4) In step (3), the eight-tube array is inserted into the handle and placed under a 300nm ultraviolet lamp for curing for 3 minutes to carry out free radical polymerization under light conditions;

[0096] (5) Take out the eight-tube array in (4) and divide it into four groups. The first group is inverted and irradiated to cure the bottom for 2 minutes and the side for 2 minutes. The second group is irradiated to cure the bottom for 2 minutes. The third group is irradiated to cure the side for 2 minutes. The fourth group is not irradiated to cure the bottom and side.

[0097] (6) Hold the handle to remove the device from the centrifuge tube and wash it three times with deionized water and anhydrous ethanol; (7) Dry at 25°C for 24 hours and then store in the dark for later use.

[0098] The comparison revealed that all four groups of nucleic acid extraction devices were intact, indicating that this experimental method does not require professional personnel, has high operational flexibility, and is suitable for rapid device construction in low-resource environments.

Claims

1. A method for preparing a hydrogel nucleic acid separation device, characterized in that, The steps are as follows: (1) Add crosslinking agent and photoinitiator to hydrogel monomer to prepare prepolymer solution; Silica particles are also added to the prepolymer solution; the silica particles used are quartz sand particles with a particle size of 50-180 mesh, and the amount of silica particles added is 20-50 wt% of the solid-liquid mixture, and the silica particles are a mixture of 50-80 mesh quartz sand and 120-180 mesh quartz sand; the crosslinking agent is one or more of N'N-methylenebisacrylamide and polyethylene glycol diacrylate, and the amount of crosslinking agent is 2-50 wt% of the hydrogel monomer; (2) After mixing the prepolymer liquid, add it to the mold, insert the handle, place it under ultraviolet light for curing, and carry out free radical polymerization reaction under light conditions to obtain the device; (3) Hold the handle to remove the device from the mold, wash and dry the device and then store it.

2. The preparation method according to claim 1, characterized in that, The hydrogel monomer is acrylic acid or acrylate.

3. The preparation method according to claim 1, characterized in that, The initiator is (2,4,6-trimethylbenzoyl)diphenylphosphine oxide, and the amount added is 0.1-0.5 wt% of the hydrogel monomer.

4. The preparation method according to claim 1, characterized in that, The free radical polymerization reaction is carried out at room temperature, with a light wavelength of 280-400 nm and an irradiation time of 3-40 min.

5. The preparation method according to claim 1, characterized in that, The mold is a cavity container that matches the needs of DNA extraction, and can be a centrifuge tube, an eight-tube array, a 96-well plate, or a deep-well plate.

6. The preparation method according to claim 1, characterized in that, The handle is rod-shaped.

7. The preparation method according to claim 1, characterized in that, The device is washed using water and / or ethanol as the washing solution, and the washing is performed at least once; the device is dried at a temperature of 0-80°C for 3 min-24 h; and the storage conditions are room temperature and protection from light.

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