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Complete-set primer applied to detecting of skin aging related susceptible genes SNP (single-nucleotide polymorphism) and application thereof

A technology for skin aging and susceptibility genes, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems of complex operation, time-consuming, incomplete gene detection, etc., to save time, The effect of reducing cost and reducing operation complexity

Inactive Publication Date: 2018-07-06
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

AI Technical Summary

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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  • Complete-set primer applied to detecting of skin aging related susceptible genes SNP (single-nucleotide polymorphism) and application thereof
  • Complete-set primer applied to detecting of skin aging related susceptible genes SNP (single-nucleotide polymorphism) and application thereof
  • Complete-set primer applied to detecting of skin aging related susceptible genes SNP (single-nucleotide polymorphism) and application thereof

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

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

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Abstract

The invention relates to a complete-set primer applied to detecting of skin aging related susceptible genes SNP (single-nucleotide polymorphism) and application thereof. The complete-set primer is prepared from amplification primer pairs of 29 SNP sites of 22 skin aging related susceptible genes of human genome DNA and extension primers; a pair of multiple PCR amplification primers and a piece ofsingle-base extension primer are designed for each site respectively, and an MALDI-TOF MS (matrix-assisted laser desorption / ionization time-of-flight mass spectrometry) method is utilized to detect SNP site genotyping of a sample to be detected. The skin aging related susceptible gene detecting method based on gene molecular subtyping disclosed by the invention can finish a plurality of SNP genotyping in the same reaction at the same time, so that accuracy, high efficiency and low cost are achieved. The gene detection disclosed by the invention can be applied to predicting of congenital agingresistance of the skin; thus, a scientific basis is provided for an external skin care scheme.

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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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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