DNA (desoxyribonucleic acid) fluorescent hydrogel and preparation method thereof

A deoxyribonucleic acid and hydrogel technology, applied in medical science, DNA/RNA fragments, bandages, etc., can solve the problems of poor bioabsorbability and degradability, and achieve the effect of reducing wound pain, soft materials, and simple process

Inactive Publication Date: 2018-01-19
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But as a medical dressing, they are poorly bioabsorbable and degradable, which can easily cause unnecessary immune responses

Method used

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  • DNA (desoxyribonucleic acid) fluorescent hydrogel and preparation method thereof
  • DNA (desoxyribonucleic acid) fluorescent hydrogel and preparation method thereof
  • DNA (desoxyribonucleic acid) fluorescent hydrogel and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] A method for preparing circular DNA with primary primers, comprising the steps of:

[0046] The specific sequences of the phosphorylated linear single-stranded DNA at the 5' end of the circular DNA with the primary primer, the primary primer, and the ssDNA added to the synthetic deoxyribonucleic acid fluorescent hydrogel are shown in Table 1.

[0047] Table 1. DNA sequences

[0048]

[0049] Wherein the nucleotide sequence of the third part in the ssDNA fragment is: 5'-ATCCTCCCACCGGGCCTCCCAC-3'SEQID NO.10

[0050] (1) Synthesis of circular DNA with primary primers:

[0051] a. DNA sample pretreatment:

[0052] Use a mini centrifuge to centrifuge the phosphorylated linear single-stranded DNA at the 5' end, the primary primer and ssDNA respectively for 1 min, throw them to the bottom of the centrifuge tube, then dissolve them in ultrapure water to 100 μM, and shake them in a constant temperature shaker for 4 hours , the condition is 30°C, 1000rpm.

[0053] b. Mix t...

Embodiment 2

[0074] A preparation method for deoxyribonucleic acid fluorescent hydrogel, comprising the steps of:

[0075] (1) Perform rolling circle amplification to obtain DNA hydrogel:

[0076] a. add the circular DNA (prepared in Example 1) with the primary primer with a final concentration of 50nmol / L according to the amount in Table 5, the phi 29 DNA polymerase with a final concentration of 0.05U / μl, and the phi 29 DNA polymerase with a final concentration of 1mmol / L dNTPs and 10×phi 29 DNA polymerase buffer and 100×BSA, NaCl solution with a final concentration of 16mmol / L, the balance is ultrapure water, the reaction condition is 200rpm shaking for 100min, and the rolling circle amplification reaction is performed to obtain long single-strand DNA.

[0077] table 5

[0078]

[0079] b. Take 100 μl of the reaction solution obtained in step a, add 2 μl of 100 μmol / L ssDNA aqueous solution, and shake at 200 rpm for 100 minutes.

[0080] The ssDNA sequence consists of three parts, ...

Embodiment 3

[0089] A method for preparing circular DNA with primary primers, comprising the steps of:

[0090] The specific sequences of the phosphorylated linear single-stranded DNA at the 5' end of the circular DNA with the primary primer, the primary primer, and the ssDNA added to the synthetic deoxyribonucleic acid fluorescent hydrogel are shown in Table 6.

[0091] Table 6. Deoxyribonucleic acid sequences

[0092]

[0093] Wherein the nucleotide sequence of the third part in the ssDNA fragment is preferred: 5'-CCCCCCCCCC-3'SEQ ID NO.11

[0094] (1) Synthesis of circular DNA with primary primers:

[0095] a. DNA sample pretreatment:

[0096] Use a mini centrifuge to centrifuge the phosphorylated linear single-stranded DNA at the 5' end, the primary primer and ssDNA respectively for 1 min, throw them to the bottom of the centrifuge tube, then dissolve them in ultrapure water to 100 μM, and shake them in a constant temperature shaker for 4 hours , the condition is 30°C, 1000rpm. ...

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Abstract

The invention discloses DNA (desoxyribonucleic acid) fluorescent hydrogel and a preparation method thereof. The preparation method comprises steps as follows: (1) DNA hydrogel is prepared; (2) the DNAhydrogel is put in sterile water, water is changed once every 15-30 min, and water is changed 3-5 times in total; (3) the DNA hydrogel obtained in the step (2) is added to a phosphate buffer and an AgNO3 water solution for a reaction; a NaBH4 water solution is added, the mixture reacts, and the DNA fluorescent hydrogel is obtained. The DNA fluorescent hydrogel has good biocompatibility and material softness, can effectively avoid secondary injury of wound as medical dressing and can relieve pain of the wound; the hydrogel has fluorescence and has certain antibacterial property and no cytotoxicity. The hydrogel preparation process is simple and convenient. The hydrogel can be applied to the field of medical dressing, immunotherapy, tissue engineering, monocellular culture, drug sustained release, protein synthesis, biosensors and the like.

Description

technical field [0001] The invention belongs to the field of deoxyribonucleic acid (DNA) material functional preparation and application, and relates to a deoxyribonucleic acid fluorescent hydrogel and a preparation method thereof. Background technique [0002] Wound dressings can protect wounds, prevent infection, and promote wound healing, and are widely used in various medical and nursing settings. At present, the materials forming hydrogel dressings are generally natural, semi-synthetic or synthetic polymers, such as: fiber derivatives (carboxymethylcellulose CMC), gelatin, polypeptides, polysaccharides, alginates, methacrylates Wait. The dressing products formed by these materials are mostly rich in water, which can provide moisture to the dry and scabbed wound surface to promote autolytic debridement; at the same time, it can absorb excess exudate from the wound surface to keep the wound surface in a proper moist state. But as a medical dressing, they are poorly bioa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L26/00C12N15/11C12P19/34
Inventor 仰大勇耿金慧寇晓虹
Owner TIANJIN UNIV
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