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Protecting method of exogenous DNA (deoxyribonucleic acid) internal standard substances in liquid products and application of exogenous DNA internal standard substances

An internal standard and product technology, applied in the field of anti-counterfeiting identification, can solve the problems of changing the physical and chemical properties of the product, difficult to apply, and high cost, and achieve the effects of low cost, convenient mass production, and easy availability of raw materials

Active Publication Date: 2017-03-15
青岛千卓分子生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because these methods need to change the physical and chemical properties of the product or add toxic and harmful chemical reagents, or require complex and expensive transformation or modification steps, it is difficult to be applied in the actual industry.

Method used

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  • Protecting method of exogenous DNA (deoxyribonucleic acid) internal standard substances in liquid products and application of exogenous DNA internal standard substances
  • Protecting method of exogenous DNA (deoxyribonucleic acid) internal standard substances in liquid products and application of exogenous DNA internal standard substances
  • Protecting method of exogenous DNA (deoxyribonucleic acid) internal standard substances in liquid products and application of exogenous DNA internal standard substances

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041]Example 1 Oligochitosan and spermine improve the tolerance of lambda DNA to DNase I

[0042] 1. Preparation of chitosan oligosaccharide and spermine solution

[0043] Preparation of chitosan oligosaccharide solution: Accurately weigh 150 mg chitosan oligosaccharide (purchased from Laizhou Haili Biological Products Co., Ltd., Shandong Province; batch number: HL130317G; molecular weight: 6500 Da; degree of deacetylation: 91.1%), add 8 mL of hydrochloric acid with pH 2.0 solution to completely dissolve the chitosan oligosaccharide, and then adjust the pH value of the solution to 7.0 with 0.1M sodium hydroxide solution. Finally, dilute to 10 mL with sterilized ultrapure water and store at 4°C. Dilute with sterilized ultrapure water to the concentration of the required working solution.

[0044] Preparation of spermine solution: Accurately weigh 100 mg of spermine (purchased from Sigma-Aldrich; product number: S3256), dissolve it in 8 mL of sterilized ultrapure water, dilut...

Embodiment 2

[0053] The protection of chitosan to polyoligonucleotide in embodiment 2 lactic acid drink

[0054] 1. Preparation of exogenous single-stranded polyoligonucleotide and chitosan complex

[0055] Preparation of chitosan solution: Accurately weigh 250 mg of chitosan (purchased from Zhejiang Aoxing Biotechnology Co., Ltd.; batch number: 20140220; molecular weight: 50-100 kDa), add 20 mL of sterilized ultrapure water, and stir magnetically for 2 h at room temperature. Fully hydrate the chitosan powder. To the hydrated chitosan solution, 20 mL of pH 2 hydrochloric acid solution was gradually added dropwise, and magnetically stirred overnight to fully dissolve it. Subsequently, its pH was adjusted to 7.0 using 0.1 M sodium hydroxide solution. Finally, dilute to 100 mL with sterilized ultrapure water and store at 4 °C. Dilute with sterilized ultrapure water to the concentration of the required working solution.

[0056] Mix the single-stranded polyoligonucleotide (36mer, the seque...

Embodiment 3

[0062] Example 3 The protection of chitosan, chitooligosaccharide and spermine in liquor to pUC18 plasmid DNA

[0063] 1. Preparation of exogenous plasmid DNA and chitosan, chitosan oligosaccharide and spermine complex

[0064] Preparation of chitosan solution: Accurately weigh 250 mg of chitosan (purchased from Sigma-Aldrich; item number: 740500; molecular weight: 110-150 kDa), add 20 mL of sterilized ultrapure water, and stir magnetically for 2 h at room temperature to make chitosan The powder hydrates well. To the hydrated chitosan solution, 20 mL of pH 2 hydrochloric acid solution was gradually added dropwise, and magnetically stirred overnight to fully dissolve it. Subsequently, the pH was adjusted to 7.0 using 0.1 M sodium hydroxide solution. Finally, dilute to 100 mL with sterilized ultrapure water and store at 4 °C. Dilute with sterilized ultrapure water to the concentration of the required working solution.

[0065] Preparation of chitosan oligosaccharide solution...

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Abstract

The invention relates to a protecting method of exogenous DNA (deoxyribonucleic acid) internal standard substances in liquid products. The protecting method especially includes steps of mixing a polycation compound solution with a purposed DNA solution according to the proportion that phosphoramidic acid ratio is not less than 10: 1, and placing for 0.5-12 hours at the temperature of 2-20 DEG C after even mixing to obtain a stable DNA compound solution which can be directly added to the liquid products, wherein the phosphoramidic acid ratio is ratio of amino moles carried in polycation to moles of phosphate groups carried in DNA. With the method, tolerance of conditions of the exogenous DNA on nuclease, acid, free radical and the like can be improved by the aid of polycation compounds, and the polycation compounds can serve as traceable and anti-counterfeiting internal standard substances existing in the liquid products or merchandises with prolonged stability.

Description

technical field [0001] The invention belongs to the technical field of anti-counterfeiting identification, and in particular relates to a method for protecting an internal standard substance of exogenous DNA in a liquid product and an application thereof. Background technique [0002] In recent years, due to the emergence of counterfeit and adulterated products, consumers pay special attention to the anti-counterfeiting and traceability of products. Especially for food products, because it is related to the health of the people, it is the focus of attention of consumers and national regulatory authorities. At present, the anti-counterfeiting and traceability measures of commodities are mainly outer packaging anti-counterfeiting, that is, attaching various special marks to the outer packaging, or using special outer packaging structures to facilitate the identification of consumers and inspection agencies. However, due to the fragility and ease of counterfeiting of the anti-...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 安然梁兴国王鹏飞
Owner 青岛千卓分子生物科技有限公司
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