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PCR (polymerase chain reaction) amplification system for genotyping of three SNP (single-nucleotide polymorphism) loci related to human folic acid metabolism and detection kit

A genotyping and human folic acid technology, applied in the field of PCR amplification systems and detection kits, can solve the problems of high cost of chip hybridization, single detection site, complicated operation, etc. Good, cost-saving effect

Active Publication Date: 2017-05-24
上海五色石医学科技有限公司
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Problems solved by technology

These methods are used in human EDTA anticoagulated whole blood MTHFR Qualitative detection of the genotype of the C677T polymorphism site, the detection site is single, and the chip hybridization method is expensive and complicated to operate

Method used

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  • PCR (polymerase chain reaction) amplification system for genotyping of three SNP (single-nucleotide polymorphism) loci related to human folic acid metabolism and detection kit
  • PCR (polymerase chain reaction) amplification system for genotyping of three SNP (single-nucleotide polymorphism) loci related to human folic acid metabolism and detection kit
  • PCR (polymerase chain reaction) amplification system for genotyping of three SNP (single-nucleotide polymorphism) loci related to human folic acid metabolism and detection kit

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Embodiment Construction

[0042] In order to better understand the content of the present invention, the specific examples of the extraction-free genotyping kit for three SNPs related to human folic acid metabolism will be further described below using a human dried blood filter paper sample. It should be understood that the following specific examples are only used to illustrate the present invention, rather than limit the present invention.

[0043] In this example, the amplification reaction was performed on an ABI 9700 thermal cycler, the electrophoresis and detection were performed on an ABI 3500 genetic analyzer, and the data analysis was performed using GeneMapper ID v3.2 software. Other reagents and materials used, such as internal standard, POP7, capillary electrophoresis buffer, and Hi-Di, are conventional materials commonly used by those skilled in the art.

[0044] : Human blood dried filter paper preparation:

[0045] Prepare a 3.0×4.0cm conventional filter paper, take 2-3 drops of periph...

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Abstract

The invention belongs to the technical field of human nucleic acid in vitro detection, and specifically relates to a PCR (polymerase chain reaction) amplification system for genotyping of three SNP (single-nucleotide polymorphism) loci related to human folic acid metabolism and a detection kit. A complex PCR amplification system comprising three SNP loci and three sample molecular marker sites is firstly designed, the three SNP loci adopt allele-specific primers, the three molecular marker sites adopt length polymorphic primers, and the primers are mutually compatible and can react in the same PCR amplification system. The detection kit comprises a primer composition container, a PCR reaction mother solution container, and a PCR auxiliary solution container, wherein the primer composition container comprises a primer SEQ ID No. 1 to SEQ ID No. 15 composition stock solution. In the case of DNA-free extraction, the three SNP loci are simultaneously amplified and typed by a PCR reaction, so that the cost, manpower and time can be obviously saved, and the working efficiency can be improved.

Description

technical field [0001] The invention belongs to the technical field of in vitro detection of human nucleic acid, and in particular relates to a PCR amplification system and a detection kit for genotyping three SNP sites related to human folic acid metabolism. Background technique [0002] One of the vitamin B complexes of folic acid, which is equivalent to pteroylglutamic acid (PGA), was extracted and purified by Mitchell (H.K. Mitchell, 1941) from spinach leaves, so it was named folic acid . The main biological function of folic acid is to participate in the methylation reaction in cells and the synthesis of deoxyribose nucleic acid as a methyl donor. Folic acid cannot be directly utilized by the human body, and must go through a series of activations to exert its physiological functions. Folic acid metabolism disorder leads to the reduction of folic acid level in the body and the increase of plasma homocysteine ​​(Hcy) concentration. The hazards related to reproductive ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6888C12Q2600/156C12Q2563/107C12Q2537/143
Inventor 赵书民赵玉莲
Owner 上海五色石医学科技有限公司
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