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Method of using taqman probe process for screening of TP53 susceptibility gene, and probe and primer used in the method

A susceptibility gene, probe method, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of high cost and time-consuming, and achieve easy interpretation and high specificity. , the effect of saving detection costs

Inactive Publication Date: 2018-05-15
美因健康科技(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the accuracy of first-generation and second-generation sequencing is relatively high, it is expensive and time-consuming

Method used

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  • Method of using taqman probe process for screening of TP53 susceptibility gene, and probe and primer used in the method
  • Method of using taqman probe process for screening of TP53 susceptibility gene, and probe and primer used in the method
  • Method of using taqman probe process for screening of TP53 susceptibility gene, and probe and primer used in the method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] A method for detecting human TP53rs78378222 gene polymorphism using Taqman MGB probe

[0039] 1. Experimental materials

[0040] Healthy people who have undergone TP53 generation sequencing were selected as test objects.

[0041] 2. Extraction of genomic DNA: Oral mucosal cells are scraped with a buccal swab, first preserved with buccal swab preservation solution, cell lysate lyses cells to release genomic DNA, and then protein precipitation solution selectively precipitates to remove protein, and finally pure genomic DNA Precipitate by isopropanol and redissolve in DNA lysate. The extraction of genomic DNA is not limited to this method, and genomic DNA extracted by other methods is also within the scope of this experiment.

[0042] 3. Genotyping was performed by Taqman MGB probe method, and the rs78378222 locus primer and probe pair were used for detection.

[0043] Primers and probes were designed for nucleotides at locus 7571752 on chromosome 17.

[0044] The nuc...

Embodiment 2

[0066] A method for detecting human TP53rs12951053 gene polymorphism using Taqman MGB probe

[0067] 1. Experimental materials

[0068] Healthy people who have undergone TP53 generation sequencing were selected as test objects.

[0069] 2. Extraction of genomic DNA: Use oral swab to scrape oral mucosal cells, first use oral swab preservation solution for preservation, use Zhiang Bio's oral swab DNA extraction kit for DNA extraction, Qubit quantification, NanoDrop detection 260 / 280 and 260 / 230 is eligible. Professionals can understand that the extraction of genomic DNA is not limited to this method, and genomic DNA extracted by other methods is also within the scope of this experiment.

[0070] 3. Genotyping was performed by Taqman MGB probe method, and the rs12951053 locus primer and probe pair were used for detection.

[0071] Primers and probes were designed for nucleotides at locus 7577407 on chromosome 17.

[0072] The nucleotide sequences of primers for the genes are ...

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Abstract

The invention provides a method of using taqman probe process for screening of TP53 susceptibility gene. The method includes the steps of: 1) extracting genome from normal people in a manner of oral cavity swab; 2) designing TP53 primers and probes; 3) performing PCR reaction to the genome with the TP53 primers and probes; 4) after the PCR reaction is finished, performing fluorescence reading andgenetic typing; 5) performing comprehensive evaluation to tumor susceptibility of individual by combining the results detected by the four probes. The taqman primers and probes have excellent genetictyping effect and can very accurately determine the genotype of the individual.

Description

technical field [0001] The invention relates to a method for tumor susceptibility gene screening and fluorescence quantitative PCR, in particular to a method for TP53 susceptibility gene screening by taqman probe method. Background technique [0002] Cancer is the biggest threat to human health in the 21st century. Although the environment and living habits have an important impact on the occurrence of cancer, cancer is essentially a genetic disease, and genetic factors increase our susceptibility to cancer. In layman's terms, single nucleotide polymorphism (SNP), which mainly refers to the DNA sequence polymorphism caused by single nucleotide variation at the genome level. It is a very ideal and effective molecular marker. [0003] The P53 protein translated by the TP53 gene is an important regulator of cell growth, proliferation and damage repair. TP53 gene is a tumor suppressor gene with low expression in normal cells and high expression in malignant tumors. It is locat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6886C12Q2600/156C12Q2561/101C12Q2531/113
Inventor 郭新雨肖哲焦少灼戚珠云郑乃中陈增辉王琦童温稳
Owner 美因健康科技(北京)有限公司
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