Compound amplification kit for simultaneously detecting female skin and hair SNP (Single Nucleotide Polymorphism) genetic polymorphism site and application thereof
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A technique of genetic polymorphism and multiple amplification, which is applied in the field of multiple amplification kits for simultaneous detection of female skin and hair SNP genetic polymorphism sites, can solve the problems of few, high cost of one-time detection, and high detection position. wait for less
Active Publication Date: 2018-05-11
AGCU SCIENTECH +1
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[0005] At present, there are not many such tests on the market. The common ones are the tests of skin anti-oxidation, anti-aging and moisturizing ability. There are few detection sites, and the detection method is chip detection. Check product, not yet available
The above detection status limits the application value of genetic testing in women's skin and hair quality
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Embodiment 1
[0097] A multiplex amplification kit for simultaneously detecting female skin and hair SNP genetic polymorphism sites, the kit contains primers for 6 SNP loci, and the 6 SNP loci are: MC1R (rs1805007), NQO1(rs1800566), AQP3(rs17553719), FGFR2(rs4752566), EDAR(rs3827760), MMP1(rs1799750); the rs number in parentheses indicates the number of the site in the dbSNP database.
[0098]The primers include shared upstream primers and specific downstream primers for each SNP locus, and the specific downstream primers introduce mismatched bases at the 2nd, 3rd or 4th position of the 3' end.
[0099] The primers and their sequences are as follows:
[0100] The shared upstream primers used to detect locus MC1R rs1805007C / T / G / A are:
[0101] SEQ1: 5'-ATGAGCACCAGCATGGCTC-3';
[0102] The specific primer used to detect site MC1R rs1805007C is one of the following nucleotide sequences:
[0103] SEQ2: 5'-TCTCCATCTTCTACGCA T TGC-3';
[0104] SEQ3: 5'-TCTCCATCTTCTACGCAC G GC-3';
[0105] ...
Embodiment 2
[0179] 1. Collection of test materials: The test materials were oral swabs of volunteers, a total of 6 copies.
[0180] 2. Template DNA extraction: The template DNA was extracted from the oral swabs of 6 volunteers by Chelex-100 method.
[0181] 3. PCR amplification:
[0182] (1), the multiple amplification detection primers used are:
[0183] MC1Rrs1805007
SEQ1: 5'-ATGAGCACCAGCATGGCTC-3'
C
SEQ4: 5'-TCTCCATCTTCTACGCACT T C-3'
T
SEQ5: 5'-atttCATCTCCATCTTCTACGCA T TGT-3'
G
SEQ9: 5'-atttcttTCTCCATCTTCTACGCAC G GG-3'
A
SEQ12: 5'-atttGGCATCTCCATCTTCTACGCACG A T-3'
NQO1rs1800566
SEQ14: 5'-GCATGGAAGCTGTTGACTTAC-3'
C
SEQ16: 5'-CAATGCTATATGTCAGTT A CGG-3'
T
SEQ20: 5'-atttCAATGCTATATGTCAGTTG A TA-3'
A
SEQ22: 5'-atttattCAATGCTATATGTCAGTTG T GT-3'
AQP3rs17553719
SEQ24: 5'-AGAGTTTGGGGAGTGCATCG-3'
T
SEQ27: 5'-ATCCGCTGCTCGCTGCT C GA-3'
G
SEQ29: 5'-atttA...
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Abstract
The invention discloses a compound amplification kit for simultaneously detecting a female skin and hair SNP (Single Nucleotide Polymorphism) genetic polymorphism site and application thereof. The kitcomprises primers of 6 SNP site genetic loci; the 6 SNP site genetic loci comprise: MC1R (rs1805007), NQO1 (rs1800566), AQP3 (rs17553719), FGFR2 (rs4752566), EDAR (rs3827760) and MMP1 (rs1799750); anrs number in brackets represents the number of the site in a dbSNP database. According to the kit disclosed by the invention, the primers are optimized so that the recognition capability of the primers to SNP is improved and the primers can be used for effectively identifying the difference of one basic group; the most suitable annealing temperature range is enlarged and a condition that a detection result has false positive, caused by the fact that annealing temperature is required to be accurate, is avoided, and the detection accuracy and reliability are improved; an ARMS (Amplification Refractory Mutation System) technology is combined with a capillary electrophoresis method for the first time; four female skin genes and 2 hair genes are simultaneously detected and the kit is a kit which has the most female skin and hair detection sites at present; the kit is convenient to use, an operation method is simple and the detection duration is shortened.
Description
technical field [0001] The invention belongs to the field of molecular biology and relates to a molecular marker and its application, in particular to a composite amplification kit for simultaneously detecting female skin and hair SNP genetic polymorphism sites and its application. Background technique [0002] Skin care and hairdressing have always been the focus of women's attention. Consumer survey research shows that as the economy grows, women spend more and more on skin care and hairdressing. However, the increasing investment in consumption does not really bring women good skin and hair quality, but often leads to more serious consequences such as roughness and dullness or hair split ends and knots. [0003] Research has shown that genetic differences in people cause skin and hair properties to vary greatly between individuals. MMP1 (matrix metalloproteinase 1), also known as interstitial collagenase, belongs to the family of matrix metalloproteinases, and its main ...
Claims
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