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DNA (deoxyribonucleic acid) extraction and shearing technology for efficiently preparing poly-desoxyribonucleic acid for skin

A polydeoxyribonucleic acid and skin technology, which is applied in the field of DNA extraction and cutting for efficient preparation of polydeoxyribonucleic acid for skin, can solve problems such as cleavage, harsh enzymes, tightness or controllability, and achieve improved yield and stability Sex, length dispersion is small, and the effect of improving product quality

Pending Publication Date: 2022-03-01
山东省循证医学研究院有限公司
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the enzyme digestion method has relatively mild reaction conditions and does not require expensive instruments, the conditions required for enzyme activity are often harsh, the amount of enzyme used is not easy to control, and the distribution of fragment sizes is not as tight or predictable as acoustic shearing. control
Sonication can produce DNA with smaller fragments, but during sonication, the sample is exposed to a large amount of energy, most of which is converted into heat, which in turn leads to strand cleavage, DNA damage and bias

Method used

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  • DNA (deoxyribonucleic acid) extraction and shearing technology for efficiently preparing poly-desoxyribonucleic acid for skin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] A preparation method of small molecule polydeoxyribonucleic acid for skin, the steps are as follows:

[0053] Step 1: Take out the testis of fresh salmon, clean the testis with pre-cooled normal saline (0.9% NaCl), and remove the blood.

[0054] Step 2: Weigh 100 g of cleaned fresh testis tissue, add 4-5 times the volume of extraction buffer (10mM Tris-HCl, 0.4mMNaCl, 2mMNa 2 EDTA), use a homogenizer to crush the testis homogenate into a paste.

[0055]Step 3: Add 4 mg of proteinase K to the crushed homogenate, and perform constant temperature enzymolysis at 37°C for 16 hours. After the enzymolysis, the mixed system is centrifuged at 10,000 rpm for 30 minutes, and the supernatant is collected.

[0056] Step 4: Add an equal volume of saturated NaCl (6M) solution to the supernatant obtained in Step 3, stir well, and centrifuge the mixed system at a speed of 10,000 rpm for 30 min to collect the supernatant and discard the precipitate.

[0057] Step 5: Add an equal volume...

Embodiment 2

[0063] To evaluate the topical efficacy of the prepared PDRN, we formulated the PDRN formulation using an acceptable cosmetic base. It includes the following raw materials and components by weight: 1-10 parts of PDRN, 5 parts of hyaluronic acid, and 100 parts of acceptable cosmetic bases (including antioxidants, moisturizers, emulsifiers, preservatives, deionized water, etc.).

[0064] The above PDRN compound solution was used to conduct wound healing experiments to evaluate the function of PDRN.

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Abstract

The invention relates to a DNA extraction and shearing technology for efficiently preparing poly-desoxyribonucleic acid for skin, which comprises the following steps: (1) taking out a salmon testis, and crushing and homogenizing by using a crusher; (2) adding protease K for enzymolysis of protein, centrifuging and collecting supernate; (3) adding a saturated sodium chloride solution, centrifuging and collecting supernate; (4) adding absolute ethyl alcohol, precipitating the fish testis DNA at low temperature, and washing the precipitate twice with 75% ethyl alcohol; (5) shearing the DNA by using a genome DNA fragmentation instrument based on an automatic sound wave focusing (TM), AFA (Automatic Focusing Acoustic Focusing) technology; and (6) freeze-drying to obtain the PDRN. According to the technology, PDRN with high yield and target molecular weight can be obtained under mild conditions. The molecular weight of the obtained PDRN ranges from 200 bp to 300 bp, and the obtained PDRN is stable in property, suitable for skin and easy to absorb. When the technology provided by the invention is used for preparing the PDRN, the molecular weight of the PDRN can be more accurately controlled, and the stability of the product is improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a DNA extraction and cutting technology for efficiently preparing polydeoxyribonucleic acid for skin. Background technique [0002] PDRN is a mixture of polydeoxyribonucleotides with a molecular weight between 50-1500KDa, which comes from the sperm DNA of salmon or rainbow trout. Numerous preclinical and clinical studies have demonstrated wound-healing and anti-inflammatory properties of PDRN, which are mediated by activation of adenosine A2A receptors and salvage pathways, in addition to promoting osteoblast activity, collagen synthesis, and angiogenesis . In fact, PDRN has been marketed for its therapeutic properties in various wound healing and inflammation-related diseases. In addition to its application in medicine, PDRN also has broad application prospects in cosmetic restoration based on its effects in promoting human cell regeneration, accelerating wound healing, reducing s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10A61K8/60A61Q19/00
CPCC12N15/1003A61K8/606A61Q19/00
Inventor 孙雯杜兆伦徐文茂
Owner 山东省循证医学研究院有限公司
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