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A complete set of primers for detecting SNPs of susceptibility genes related to skin aging and its application

A technology for skin aging and susceptibility genes, which is applied in the direction of recombinant DNA technology, biochemical equipment and methods, and microbial measurement/inspection. The effect of reducing cost and reducing operation complexity

Inactive Publication Date: 2021-08-13
北京天平永达生物科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Invention patent (201610367682.9) discloses a primer and probe for detecting skin water retention. The method used is the method of fluorescent quantitative PCR. There are two single nucleotide polymorphism sites of the detected gene, and the detected gene is not Comprehensive, and the fluorescent quantitative PCR method is suitable for detecting about 5 known SNP sites. When the number of sites increases, the operation is complicated and time-consuming

Method used

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  • A complete set of primers for detecting SNPs of susceptibility genes related to skin aging and its application
  • A complete set of primers for detecting SNPs of susceptibility genes related to skin aging and its application
  • A complete set of primers for detecting SNPs of susceptibility genes related to skin aging and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] The establishment of embodiment 1 technical scheme

[0054] S1 According to the relationship between human skin aging and genetic genes, determine skin aging-related susceptibility genes and SNP sites (Table 1).

[0055] S2 Design a set of primers for each SNP site, including a pair of amplification primers (Table 2) and a single base extension primer (Table 3).

[0056] S3 uses human genomic DNA as a sample, and a set of primers is amplified by multiplex PCR.

[0057] S4 uses matrix-assisted laser desorption ionization time-of-flight mass spectrometry to detect the SNP site type of the sample, and predict the skin anti-aging ability of the sample.

[0058] Table 1 Susceptibility genes and SNP sites related to skin aging

[0059]

[0060]

[0061] Table 2 Multiplex PCR amplification primer sequence

[0062]

[0063]

[0064] Table 3 Single base extension primer sequence

[0065] site serial number extension primer sequence rs10733310 59...

Embodiment 2

[0066] Example 2 sample extraction and analysis

[0067] In order to verify the detection accuracy and effectiveness of the set of primers used to detect the susceptibility gene SNP associated with skin aging of the present invention, 4 samples were selected for genetic detection, and the process was as follows:

[0068] A. Saliva DNA extraction: use Kangwei Century Oral Swab Genomic DNA Extraction Kit for saliva DNA extraction. Add a certain amount of absolute ethanol to the GW1 and GW2 reagents. Take 500 μL saliva sample from the sampling tube, add 300 μL GR, 20 μL Proteinase K and 300 μL GL, shake and mix, incubate with shaking at 56°C for 15 minutes, add 300 μL absolute ethanol, shake and mix; add 750 μL of this solution to the adsorption tube of the collection tube Add 400 μL GW1 to the adsorption column, centrifuge at 12,000 rpm for 1 min, discard the liquid; add 400 μL GW2 to the adsorption column, centrifuge at 12,000 rpm for 1 min, discard the liquid; centrifuge at 1...

Embodiment 3

[0074] Example 3 Analysis of genetic testing results

[0075] The 29 SNP site gene detection results (Table 5) of the 4 samples in Example 2 were analyzed, combined with the interpretation of the significance of the SNP site typing of skin aging-related susceptibility genes (Table 6), the skin of the tested sample Analysis of anti-aging ability.

[0076] Table 5 SNP site gene detection results

[0077] site sample 1 sample 2 sample 3 Sample 4 rs10733310 GT GT GT G rs62543565 C C C C rs1015362 CT C CT C rs4911414 GT G GT G rs322458 CT C C C rs1540771 C C C C rs1799750 C C C C rs2070600 C C C C rs2071288 C C C C rs1130534 T T T T rs16891982 C C C C rs1834640 G G G G rs642742 TC C CT C rs1042602 C C C C rs2031526 AG AG G AG rs885479 GA GA G G rs74653330 C C C C rs1800414 C CT CT CT rs4880 A A A A...

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Abstract

The present invention relates to a set of primers for detecting SNPs of susceptibility genes related to skin aging and the application thereof. The set of primers includes a pair of amplification primers for 29 SNP sites of 22 skin aging-related susceptibility genes of human genomic DNA and extension primers. A pair of multiplex PCR amplification primers and a single-base extension primer were designed for each site, and MALDI-TOF MS (matrix-assisted laser desorption ionization time-of-flight mass spectrometry) method was used to detect the SNP site typing of the samples to be tested. The invention establishes a detection method for skin aging-related susceptibility genes based on gene molecular typing, which can complete multiple SNP typing in the same reaction at the same time, which is accurate, efficient and low in cost. The gene detection of the present invention is used to predict the innate anti-aging ability of the skin, thereby providing a scientific basis for external skin care regimens.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a set of primers for detecting SNPs of susceptibility genes related to skin aging and applications thereof. Background technique [0002] The Dermatologist Branch of the Chinese Medical Doctor Association launched a "large-scale online survey on the skin condition of Chinese people". Nearly 40,000 Internet users participated in the survey, of which 45.45% were men and 54.55% were women. The age group of 29 to 39 has the largest number of people, accounting for 39.45%. The results show that about 70% of people have sub-healthy skin, and only 14% of people have good skin condition. Human skin over the age of 25 begins to enter the process of natural aging. Skin aging mainly focuses on three aspects: texture, color, and fineness. Specifically, on the face, skin aging can be manifested as skin relaxation, lack of elasticity, wrinkles, dull color, Enlarged pores, dilated capil...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 张影
Owner 北京天平永达生物科技发展有限公司
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