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Primer for detecting folic acid metabolism related gene SNP, fluorescent probe and application

A folic acid metabolism and fluorescent probe technology, applied in the biological field, can solve the problems of insufficient, cross-contaminated PCR products, enzyme digestion, low reliability, etc., and achieve the effects of low cost, accurate and reliable detection results, and good practicability

Inactive Publication Date: 2014-10-22
湖北维达健基因技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method is complicated to operate, and when the sample size is large, it is easy to cause cross-contamination of PCR products, and it is prone to insufficient or excessive digestion, resulting in false negative or false positive results, and low reliability.
Although the specificity of the multiplex PCR method has been improved, the principle of this method is still based on the principle of ordinary PCR, and factors such as primer specificity and low-fidelity Taq enzymes can affect the results
Although the DNA sequencing method is currently the gold standard for genetic diagnosis, the steps are cumbersome, the process is complicated, the reagents are expensive, and cross-contamination between samples is prone to occur, resulting in sequencing failures; in addition, the equipment of the sequencer is beyond that of a general clinical laboratory. Tolerance range
The high-resolution melting curve method is a fast, simple, economical and practical typing method, but genotyping depends on the precision of the temperature control of the instrument, and the false positive is high.

Method used

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  • Primer for detecting folic acid metabolism related gene SNP, fluorescent probe and application
  • Primer for detecting folic acid metabolism related gene SNP, fluorescent probe and application
  • Primer for detecting folic acid metabolism related gene SNP, fluorescent probe and application

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Experimental program
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Embodiment

[0087] 1. Prepare the following primers and probes:

[0088] (1) Primers and fluorescent probes for the rs1801133 SNP site of the methylenetetrahydrofolate reductase gene:

[0089] Forward primer MTHFR-F: 5'-TACCCCAAAGGCCACCCCG-3'

[0090] Reverse primer MTHFR-R: 5'-CATGCCTTCACAAAGCGGA-3'

[0091] MTHFR-T probe:

[0092] FAM-GTGTCTGCGGGAG TCGATTTCATCATCACGC-MGB

[0093] MTHFR-C probe:

[0094] VIC-GTGTCTGCGGGAG C CGATTTCATCATCACGC-MGB

[0095] (2) Primers and fluorescent probes for the rs11018628 SNP site of the NOX oxidase gene:

[0096] Forward primer NOX-F: 5'-TGCAGAGAGCATGGTTGACTAG-3'

[0097] Reverse primer NOX-R: 5'-TAAAGAGGAATGAGGCAACAT-3'

[0098] NOX-T probe:

[0099] FAM-TGATTGTTTTTGATG T GGAGTAGATGAGAG-MGB

[0100] NOX-C probe:

[0101] VIC-TGATTGTTTTTGATG C GGAGTAGATGAGAG-MGB

[0102] (3) Primers and fluorescent probes for the rs1047891 SNP site of the carbamyl phosphate synthase gene:

[0103] Forward primer CPS-F: 5'-TAAATTATTTCAGAAAACAG-3'

[01...

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Abstract

The invention discloses primers for detecting folic acid metabolism related gene SNP, fluorescent probes and applications. The provided primers have four groups. Each group of the primers has a corresponding fluorescent probe group for carrying out rapid gene typing on rs1801133, rs11018628 and rs1047891. Detected results obtained by the primers and the fluorescent probe are accurate and reliable. The primers have low cost and relatively good practicality.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer, a fluorescent probe and an application for detecting the SNP of a gene related to folic acid metabolism. Background technique [0002] Folic acid is one of the vitamin B complexes, which is equivalent to pteroylglutamic acid (PGA). It was extracted and purified by Mitchell (H.K. Mitchell, 1941) from spinach leaves, so it is named folic acid. Folic acid is an important nutrient needed by the body, and it is rich in green leafy vegetables, fruits and animal liver. Folic acid works in the human body in the form of tetrahydrofolate. Tetrahydrofolate is a carrier of one-carbon units, which can carry one-carbon units such as methyl, methylene, and formaldehyde, and participate in the synthesis of amino acids, purines, and pyrimidines in the body. Therefore, folic acid is an indispensable nutrient for cell division and proliferation, tissue repair and body development. [0003] ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6876C12Q1/6851C12Q1/6858C12Q2600/156
Inventor 徐谋胜袁雅燕黄淑君余亚军王珊
Owner 湖北维达健基因技术有限公司
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