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Method for detecting genotype of genes related to cell cycle control

A cycle regulation and cell cyclin technology, applied in the field of molecular biology, can solve the problems of false positive phenomena that cannot be well checked and explained, and achieve the effect of strong repeatability and high detection accuracy

Active Publication Date: 2011-11-09
上海中优医药高科技股份有限公司
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AI Technical Summary

Problems solved by technology

False positives in the detection of CCND1 and STK15 genes in previous detection methods cannot be well investigated and explained

Method used

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  • Method for detecting genotype of genes related to cell cycle control
  • Method for detecting genotype of genes related to cell cycle control
  • Method for detecting genotype of genes related to cell cycle control

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Experimental program
Comparison scheme
Effect test

Embodiment

[0035] A genotype detection method for genes related to cell cycle regulation:

[0036] Step 1. Type of sample tested and sampling method:

[0037] Use the sampler to wipe the left and right cheeks of the testee’s mouth 20 times to collect oral mucosal epithelial cell samples;

[0038] Step 2. Extraction method of genomic DNA:

[0039] The genomic DNA of each subject’s oral epithelial cell sample is extracted by the silica gel adsorption method for 2 hours to 2.5 hours. After electrophoresis detection, a clear white band is visible to the naked eye and the next step is to be tested;

[0040] Step 3. Fluorescence quantitative PCR reaction:

[0041] Put each test subject's sample into 2 reaction wells, and detect 2 sites at the same time, namely cyclin (CCND1) rs9344, serine / threonine kinase 15 (STK15) s2273535, and add 2 more without DNA NTC blank control hole of the template;

[0042] Adding reagents to each reaction well is a fluorescent quantitative PCR reaction system, the total volum...

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Abstract

The invention provides a method for detecting genotype of genes related to cell cycle control, which is characterized by including the following concrete steps: collecting the oral mucosa sample of the detected; extracting genomic DNA, carrying out genotyping and analyzing the result. The DNA to be detected in the invention can be obtained by extracting the oral mucosa epithelial cell of the detected and the sample collection can be carried out by employing oral mucosa epithelial cell collection method and related equipment. As far as the DNA extraction of the sample is concerned, the silica gel adsorption method can be employed to extract the DNA of the oral epithelial cell and measure purity and concentration of the extracted DNA. Two-locus Genotyping can be carried out by adopting the TaqMan-MGB technology. As proven by the experiment, accuracy rate of genotyping reaches more than 99%, while the repeatability thereof 100%.

Description

Technical field [0001] The invention relates to a method for detecting the genotype of genes related to cell cycle regulation, which is used for detecting the mutation of CCND1 and STK15 genes, and belongs to the technical field of molecular biology. Background technique [0002] A large-scale study of the Chinese population confirmed that mutations in the cyclin CCND1 and serine / threonine kinase STK15 genes are common in the Chinese population. The false positives in CCND1 and STK15 genes detected by previous detection methods cannot be investigated and explained well. Summary of the invention [0003] The purpose of the present invention is to provide a method for detecting the genotype of genes related to cell cycle regulation with high accuracy. [0004] In order to achieve the above objective, the technical solution of the present invention is to provide a genotype detection method of cell cycle regulation-related genes, which is characterized in that the specific steps are: ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 王校傅咏南
Owner 上海中优医药高科技股份有限公司
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