CN112275226A - Preparation method of microcapsule for DNA information storage - Google Patents
Preparation method of microcapsule for DNA information storage Download PDFInfo
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- 239000003094 microcapsule Substances 0.000 title claims abstract description 17
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- 108020004414 DNA Proteins 0.000 claims abstract description 64
- 239000004005 microsphere Substances 0.000 claims abstract description 49
- 229920002873 Polyethylenimine Polymers 0.000 claims abstract description 46
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 22
- 238000002844 melting Methods 0.000 claims abstract description 20
- 238000002791 soaking Methods 0.000 claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 19
- 230000008018 melting Effects 0.000 claims abstract description 12
- 239000008367 deionised water Substances 0.000 claims abstract description 11
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 238000001338 self-assembly Methods 0.000 claims abstract description 7
- 238000004140 cleaning Methods 0.000 claims abstract description 6
- 238000005119 centrifugation Methods 0.000 claims description 21
- 239000006228 supernatant Substances 0.000 claims description 19
- 229910052681 coesite Inorganic materials 0.000 claims description 18
- 229910052906 cristobalite Inorganic materials 0.000 claims description 18
- 239000000377 silicon dioxide Substances 0.000 claims description 18
- 229910052682 stishovite Inorganic materials 0.000 claims description 18
- 229910052905 tridymite Inorganic materials 0.000 claims description 18
- DDFHBQSCUXNBSA-UHFFFAOYSA-N 5-(5-carboxythiophen-2-yl)thiophene-2-carboxylic acid Chemical compound S1C(C(=O)O)=CC=C1C1=CC=C(C(O)=O)S1 DDFHBQSCUXNBSA-UHFFFAOYSA-N 0.000 claims description 9
- MIMUSZHMZBJBPO-UHFFFAOYSA-N 6-methoxy-8-nitroquinoline Chemical compound N1=CC=CC2=CC(OC)=CC([N+]([O-])=O)=C21 MIMUSZHMZBJBPO-UHFFFAOYSA-N 0.000 claims description 9
- 238000002156 mixing Methods 0.000 claims description 9
- 239000011248 coating agent Substances 0.000 claims description 3
- 238000000576 coating method Methods 0.000 claims description 3
- 239000002245 particle Substances 0.000 claims description 3
- 102000053602 DNA Human genes 0.000 abstract description 49
- 239000002775 capsule Substances 0.000 abstract description 5
- 230000009881 electrostatic interaction Effects 0.000 abstract 1
- 239000000463 material Substances 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 55
- 238000005406 washing Methods 0.000 description 9
- 239000007984 Tris EDTA buffer Substances 0.000 description 5
- 238000013500 data storage Methods 0.000 description 5
- 238000007654 immersion Methods 0.000 description 4
- XZWYZXLIPXDOLR-UHFFFAOYSA-N metformin Chemical compound CN(C)C(=N)NC(N)=N XZWYZXLIPXDOLR-UHFFFAOYSA-N 0.000 description 4
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 2
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 2
- ASJSAQIRZKANQN-CRCLSJGQSA-N 2-deoxy-D-ribose Chemical compound OC[C@@H](O)[C@@H](O)CC=O ASJSAQIRZKANQN-CRCLSJGQSA-N 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- 229930024421 Adenine Natural products 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 230000004308 accommodation Effects 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- 229940104302 cytosine Drugs 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 229940113082 thymine Drugs 0.000 description 1
Classifications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
- B01J13/02—Making microcapsules or microballoons
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
-
- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06F—ELECTRIC DIGITAL DATA PROCESSING
- G06F16/00—Information retrieval; Database structures therefor; File system structures therefor
- G06F16/90—Details of database functions independent of the retrieved data types
- G06F16/901—Indexing; Data structures therefor; Storage structures
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- Genetics & Genomics (AREA)
- General Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Databases & Information Systems (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
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- Plant Pathology (AREA)
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Abstract
The invention discloses a preparation method of microcapsules for DNA information storage, which belongs to the technical field of information storage, and comprises the steps of firstly soaking silica microspheres in a polyethyleneimine solution with positive charges, then centrifugally cleaning, then soaking the silica microspheres in a deoxyribonucleic acid solution with negative charges for a period of time, centrifugally cleaning with deionized water, repeatedly repeating the operation, assembling PEI and DNA on the surfaces of the silica microspheres for multiple times, finally soaking the microspheres subjected to layer-by-layer self-assembly in a core melting solution, and dissolving cores of the silica microspheres. Since the positively charged PEI solution and the negatively charged DNA solution can be adsorbed by electrostatic interaction, the capsules can be prepared by a layer-by-layer self-assembly method. The method is simple and convenient to operate, the materials are cheap and easy to obtain, and the preparation time is short.
Description
Technical Field
The invention belongs to the technical field of information storage, and particularly relates to a preparation method of a microcapsule for DNA information storage.
Background
With the rapid development of electronic technology and internet, the resulting explosively increasing digitally stored data causes a great deal of pressure and challenge to the data storage method, and the capacity of the existing storage medium cannot keep up with the increasing speed of the digital information, so that the traditional data storage mode is in urgent need of improvement and breakthrough, and the research of an emerging data storage technology is significant.
Deoxyribonucleic acid (DNA) biomolecules, which consist of deoxyribose, phosphate, and bases (adenine, thymine, cytosine, or guanine), are an important biological information storage medium. The DNA is used for data storage, so that the data storage is efficient, stable and resource-saving.
The storage of DNA is expected to become a new generation of storage technology, and the digital storage technology of DNA is a technology for storing target information required to be stored by utilizing an artificially synthesized deoxynucleotide chain and reading the stored information by utilizing a sequencing method.
DNA, as an emerging information storage medium, was first since 1988, and has the following advantages: breaks through the limitation of the traditional solid medium (hard disk, optical disk and mobile magnetic disk); the 0-1 coding is changed into the AGTC combined coding, a set of technical system for code storage and data reading can be formed through a unique arrangement and combination mode, and the AGTC combined coding has super-strong data accommodation and high-density data volume; the DNA can be stored for one million years at the temperature of-18 ℃, can be stored for a long time and has low subsequent maintenance cost.
Disclosure of Invention
The purpose of the invention is as follows: in order to solve the problems in the prior art, the preparation method of the microcapsule for storing the DNA information ensures the stable and accurate performance of stored information and outstanding storage density.
The technical scheme is as follows: in order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of microcapsules for DNA information storage comprises the following steps: mixing SiO2The microspheres are uniformly dispersed in the solution, and the polyethyleneimine PEI solution and the deoxyribonucleic acid DNA solution are sequentially assembled to SiO layer by using a layer-by-layer self-assembly method2Obtaining SiO with DNA coated with deoxyribonucleic acid and PEI coated with polyethyleneimine on the inner layer on the surface of the microsphere2And (3) soaking the microspheres in a core melting solution, and melting the cores of the microspheres to obtain the DNA information storage microcapsule.
Furthermore, the outer surface is coated with DNA and the inner layer is coated with SiO of PEI2The preparation method of the microsphere comprises the following steps:
step 1, SiO2Soaking the microspheres in a polyethyleneimine PEI solution with positive charges; centrifuging to remove supernatant, and washing with deionized water for multiple times to obtain SiO coated with polyethyleneimine PEI2Microspheres;
step 2: then coating the surface of the SiO coated with polyethyleneimine PEI in the step 12Soaking the microspheres in DNA solution with negative charges, centrifuging, sucking off supernatant, and repeatedly cleaning to obtain SiO coated with DNA and polyethyleneimine PEI2Microspheres; repeating the operation processes of the step 1 and the step 2, thereby obtaining SiO which is self-assembled layer by layer and adsorbs polyethyleneimine PEI and DNA layer by layer through electrostatic action2And (3) microspheres.
Further, in the step 1-step 2, the centrifugation time is 10-15 min, and the centrifugation speed is 8000-9000 rpm.
Further, the preparation method of the core-melting solution comprises the following steps: respectively dissolving ammonium bifluoride and ammonium fluoride in deionized water, wherein the concentration of the ammonium bifluoride solution is 40-50 mg/mL, the concentration of the ammonium fluoride solution is 40-42 mg/mL, and mixing the prepared ammonium bifluoride solution and the ammonium fluoride solution in equal volume to obtain a core-melting solution, wherein the pH value is 4.
Further, the SiO2The particle size of the microspheres is 1-5 μm; the concentration of the polyethyleneimine PEI solution is 1-10 mg/mL.
The invention principle is as follows: the PEI solution is electronegative and the DNA solution is electropositive, so that the PEI solution can be assembled layer by utilizing the electrostatic action of positive and negative charges, the DNA is used as a genetic information carrier, the accurate heredity of life can be ensured, and the formed DNA information storage capsule has good information storage density, so that the DNA information storage capsule prepared by the layer-by-layer self-assembly method has the performance of ensuring the stability and accuracy of stored information and outstanding storage density.
Has the advantages that: compared with the prior art, the preparation method of the microcapsule for storing the DNA information is simple and effective, is simple and convenient to operate, requires short time and simple required equipment; the DNA is used as a genetic information storage carrier, so that the preparation of life can be ensured to be inherited without error, and 2.15 hundred million G of information can be stored in 1G of DNA, so that the capsule has high information storage accuracy and large information unit storage capacity; the samples of over ten thousand years can be recovered into complete DNA fragments, so that the information storage capsule has long shelf life and is not easily interfered and inhibited by the working environment.
Detailed Description
The invention is further illustrated by the following examples.
The preparation method of the microcapsule for storing the DNA information comprises the following steps: mixing SiO2The microspheres are uniformly dispersed in the solution, and the polyethyleneimine PEI solution and the deoxyribonucleic acid DNA solution are sequentially assembled to SiO layer by using a layer-by-layer self-assembly method2Obtaining SiO with DNA coated with deoxyribonucleic acid and PEI coated with polyethyleneimine on the inner layer on the surface of the microsphere2Then soaking the obtained microsphere in a core-melting solution to melt SiO2The micro-capsule for storing DNA information is obtained from the micro-capsule inner core.
SiO coated with DNA and PEI2The preparation method of the microsphere comprises the following steps:
step 1, SiO2Soaking the microspheres in a polyethyleneimine PEI solution with positive charges; centrifuging to remove supernatant, and washing with deionized water for multiple times to obtain SiO coated with polyethyleneimine PEI2Microspheres;
step 2: then coating the surface of the SiO coated with polyethyleneimine PEI in the step 12Soaking the microspheres in DNA solution with negative charges, centrifuging, removing supernatant, and repeatedly cleaning for several times to obtain SiO coated with DNA and PEI2Microspheres; repeating the operation processes of the step 1 and the step 2, thereby obtaining SiO which adsorbs PEI and DNA layer by layer through electrostatic action after self-assembly layer by layer2And (3) microspheres.
In the step 1-step 2, the centrifugation time is 10-15 min, and the centrifugation speed is about 8000-9000 rpm.
The preparation method of the core melting solution comprises the following steps: respectively dissolving ammonium bifluoride and ammonium fluoride in deionized water, wherein the concentration of the ammonium bifluoride solution is 40-50 mg/mL, the concentration of the ammonium fluoride solution is 40-42 mg/mL, equivalently mixing the prepared ammonium bifluoride solution and the prepared ammonium fluoride solution to obtain a core-melting solution, and adjusting the pH to 4.
Specifically, SiO2Even dispersion of the microspheres by oscillation, SiO2The particle size of the microspheres is about 5 μm. Then SiO2Soaking the microspheres in a positively charged polyethyleneimine PEI solution for about 8-10 min, wherein the concentration of the PEI solution is 1 mg/mL; after centrifuging to remove supernatant, washing with deionized water for multiple times (3-5 times). After further immersion in a negatively charged DNA solution for a certain period of time, the DNA powder was dissolved in TE buffer at a concentration of 1.8 mM. Similarly, soaking for 8-10 min, centrifuging and removing supernatant, wherein the centrifugation speed and the centrifugation time are the same as above. Repeating the above process for multiple times to obtain SiO for adsorbing PEI and DNA layer by layer through electrostatic action2The microspheres are finally soaked in a core melting solution to melt SiO2The microspheres were cored until the solution was clear and transparent, and after centrifugation, excess core-melting solution was washed away with TE buffer.
Example 1:
mixing SiO2The microspheres are oscillated and dispersed uniformly, and then SiO is added2Soaking the microspheres in a positively charged PEI solution for about 8-10 min, wherein the concentration of the PEI solution is 1 mg/mL; after centrifuging to remove the supernatant, washing the supernatant for multiple times (3-5 times) by using deionized water, wherein the centrifugation speed is 8000-9000 rpm, and the centrifugation time is 10-12 min. After further immersion in a negatively charged DNA solution for a period of time, the DNA concentration was 1.8 mM. Similarly, soaking for 8-10 min, centrifuging and removing supernatant, wherein the centrifugation speed and the centrifugation time are the same as above. Repeating the above process for multiple times to obtain SiO for adsorbing PEI and DNA layer by layer2The microspheres are finally soaked in 10mL of core melting solution to melt SiO2And (4) carrying out core treatment on the microspheres until the solution is clear and transparent, centrifuging, removing the supernatant, and washing the excessive core melting solution by using a TE buffer solution.
Example 2:
mixing SiO2The microspheres are oscillated and dispersed uniformly, and then SiO is added2Soaking the microspheres in a positively charged PEI solution for about 4-6 min, wherein the concentration of the PEI solution is 0.5 mg/mL; and after centrifuging to remove the supernatant, washing the supernatant for multiple times (3-5 times) by using deionized water, wherein the centrifugation speed is 6000-7000 rpm, and the centrifugation time is 8-10 min. After further immersion in a negatively charged DNA solution for a period of time, the DNA concentration was 0.9 mM. Similarly, soaking for 5-7 min, centrifuging and removing supernatant, wherein the centrifugation speed and the centrifugation time are the same as above. Repeating the above process for multiple times to obtain SiO for adsorbing PEI and DNA layer by layer2The microspheres are finally soaked in 10mL of core melting solution to melt SiO2And (4) carrying out core treatment on the microspheres until the solution is clear and transparent, centrifuging to remove supernatant, and washing away excessive core melting solution by using TE buffer solution.
Example 3:
mixing SiO2The microspheres are oscillated and dispersed uniformly, and then SiO is added2Soaking the microspheres in a positively charged PEI solution for about 12-15 min, wherein the concentration of the PEI solution is 2 mg/mL; centrifuging to remove supernatant, and then washing with deionized water for multiple times (5-7 times), wherein the centrifugation speed is 10000-11000 rpm, and the centrifugation time is 12-14 min. After further immersion in a negatively charged DNA solution for a period of time, the DNA concentration was 3.6 mM. Similarly, soaking for 12-15 min, then centrifuging and sucking out the supernatant, wherein the centrifugation speed and the centrifugation time are the same as above. Repeating the above process for multiple times to obtain SiO for adsorbing PEI and DNA layer by layer2The microspheres are finally soaked in 15mL of core-melting solution to melt SiO2And (4) carrying out inner core treatment on the microspheres until the solution is clear and transparent, centrifuging to remove supernatant, and washing away excessive core melting solution by using a TE buffer solution.
Claims (5)
1. A preparation method of microcapsules for DNA information storage is characterized by comprising the following steps: mixing SiO2The microspheres are uniformly dispersed in the solution, and the polyethyleneimine solution and the deoxyribonucleic acid DNA solution are sequentially assembled to SiO layer by using a layer-by-layer self-assembly method2The surface of the microsphere is coated with DNA and the inner layer is coated with polyethyleneimine2And (3) soaking the microspheres in a core melting solution, and melting the cores of the microspheres to obtain the DNA information storage microcapsule.
2. The method according to claim 1, wherein said DNA information storage microcapsule is coated with DNA and an inner layer of SiO coated with polyethyleneimine2The preparation method of the microsphere comprises the following steps:
step 1, SiO2Soaking the microspheres in a polyethyleneimine solution with positive charges; centrifuging to remove supernatant, and cleaning with deionized water to obtain SiO coated with polyethyleneimine2Microspheres;
step 2: then coating the surface of the step 1 with SiO of polyethyleneimine2Soaking the microspheres in DNA solution with negative charges, centrifuging, removing supernatant, and repeatedly cleaning to obtain SiO coated with DNA and polyethyleneimine2Microspheres; repeating the operation processes of the step 1 and the step 2, thereby obtaining SiO which is self-assembled layer by layer and adsorbs polyethyleneimine and DNA layer by layer through electrostatic action2And (3) microspheres.
3. The method for preparing microcapsules for storing DNA information as claimed in claim 2, wherein the centrifugation time is 10 to 15min and the centrifugation speed is 8000 to 9000rpm in the steps 1 to 2.
4. The method for preparing microcapsules for DNA information storage according to claim 1, wherein the method for preparing the core-melting solution is as follows: respectively dissolving ammonium bifluoride and ammonium fluoride in deionized water, wherein the concentration of the ammonium bifluoride solution is 40-50 mg/mL, the concentration of the ammonium fluoride solution is 40-42 mg/mL, and isovolumetrically mixing the prepared ammonium bifluoride solution and the prepared ammonium fluoride solution to obtain a core-melting solution, wherein the pH value is 4.
5. The method for preparing microcapsules for DNA information storage according to claim 1, wherein said SiO is2The particle size of the microspheres is 1-5 μm; the concentration of the polyethyleneimine solution is 1-10 mg/mL.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2023172577A1 (en) * | 2022-03-07 | 2023-09-14 | Core Laboratories Lp | Oligonucleotide-containing tracer particles for subterranean applications |
| US11760925B1 (en) | 2022-03-07 | 2023-09-19 | Core Laboratories Lp | Oligonucleotide-containing tracer particles for subterranean applications |
| CN116905093A (en) * | 2023-06-07 | 2023-10-20 | 祥符实验室 | Long-term preservation method of DNA database and application thereof |
| CN116905093B (en) * | 2023-06-07 | 2024-05-24 | 祥符实验室 | Long-term preservation method of DNA database and application thereof |
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| CN107119111A (en) * | 2017-03-28 | 2017-09-01 | 浙江大学 | A kind of DNA molecular LBL self-assembly microcapsules of DNA enzymatic parcel and its preparation and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107119111A (en) * | 2017-03-28 | 2017-09-01 | 浙江大学 | A kind of DNA molecular LBL self-assembly microcapsules of DNA enzymatic parcel and its preparation and application |
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| YAN-ZHI TIAN ET AL: "Nuclease-responsive DNA–PEI hollow microcapsules for bio-stimuli controlled release", 《J. MATER. CHEM. B》 * |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023172577A1 (en) * | 2022-03-07 | 2023-09-14 | Core Laboratories Lp | Oligonucleotide-containing tracer particles for subterranean applications |
| US11760925B1 (en) | 2022-03-07 | 2023-09-19 | Core Laboratories Lp | Oligonucleotide-containing tracer particles for subterranean applications |
| CN116905093A (en) * | 2023-06-07 | 2023-10-20 | 祥符实验室 | Long-term preservation method of DNA database and application thereof |
| CN116905093B (en) * | 2023-06-07 | 2024-05-24 | 祥符实验室 | Long-term preservation method of DNA database and application thereof |
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