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Multiplex real-time fluorescent PCR detection primers and probes for TP53 gene SNP loci and multiplex real-time fluorescent PCR detection kit

A real-time fluorescent and fluorescent probe technology, applied in DNA/RNA fragments, recombinant DNA technology, microorganism determination/inspection, etc., can solve the problems of high cost and low detection throughput, and achieve low cost, high sensitivity, method simple effect

Inactive Publication Date: 2017-11-21
厦门智因互联科技有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, the common method used for genetic screening is Sanger sequencing, which can sequence a certain region of a sample, but the detection throughput is low and the cost is high

Method used

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  • Multiplex real-time fluorescent PCR detection primers and probes for TP53 gene SNP loci and multiplex real-time fluorescent PCR detection kit
  • Multiplex real-time fluorescent PCR detection primers and probes for TP53 gene SNP loci and multiplex real-time fluorescent PCR detection kit
  • Multiplex real-time fluorescent PCR detection primers and probes for TP53 gene SNP loci and multiplex real-time fluorescent PCR detection kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: Accuracy detection experiment

[0038] Extraction of DNA from the sample to be tested: Genomic DNA is extracted from the oral epithelial cells of the subject using conventional molecular biology methods or commercially available kits.

[0039] Real-time fluorescent PCR amplification and detection:

[0040] The reaction system is: 10mM Tris-HCl (pH8.3), 50mM KCl, 2mM MgCl 2 , upstream primers AC1-F, AC2-F each 10pmol, downstream primers AC1-R, AC2-R each 10pmol and FAM-labeled fluorescent probe AC1 C-P, HEX-labeled fluorescent probe AC1 A-P, ROX-labeled fluorescent probe AC2 A-P, CY5-labeled fluorescent probe AC2 G-P 5pmol each, sample DNA template 20ng; water to make up to 25μL.

[0041] Where AC1: rs12951053 site

[0042]

[0043] AC2: rs2078486 locus

[0044]

[0045] The real-time fluorescent PCR reaction was carried out on the Bio-RadCFX96 instrument, and the amplification detection was carried out according to the following conditions:

[0...

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Abstract

The invention discloses multiplex real-time fluorescent PCR detection primers and probes for TP53 gene SNP loci and multiplex real-time fluorescent PCR detection kit. The primers are upstream primers AC1-F and AC1-R, AC2-F and AC2-R; the probes are AC1 C-P, AC1 A-P, AC2 A-P and AC2 G-P; the 5 'ends of the probes are labeled with fluorescent groups and the 3' ends are labeled with quenching groups, wherein the nucleotide sequences of AC1-F, AC1-R, AC2-F, AC2-R, AC1 C-P, AC1 A-P, AC2 A-P and AC2 G-P are as shown in SEQ ID NO: 1-8 respectively. The primers, the probes and the kit are used for detecting the TP53 gene SNP loci, and have the advantages of simple method, accuracy, high sensitivity, low cost, high flux and suitability for general screening.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a multiplex real-time fluorescent PCR detection primer, probe and kit for SNP sites of TP53 gene. Background technique [0002] Malignant tumors have become the most important factor threatening the health of Chinese people and the number one cause of death. From the perspective of incidence rate, the top five cancers in men are lung cancer, liver cancer, colon cancer, stomach cancer, and genitourinary system cancer, and the top five cancers in women are breast cancer, lung cancer, reproductive system cancer, bowel cancer, and stomach cancer. [0003] TP53 is a tumor suppressor gene. This gene has a protective effect on the human body. It monitors the integrity of the gene at all times. Once the DNA of the cell is damaged, it can prevent the cell from entering the DNA synthesis period, inhibit the division and proliferation of the cell, and repair the damaged DNA, even if it cannot ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/156C12Q2600/16
Inventor 朱莎莎叶国栋陈茂立许剑雄徐晓政董康梅陈巍魏李彬李顺杰
Owner 厦门智因互联科技有限公司
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