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TERT gene polymorphism detection kit through pyrosequencing method, and method thereof

A technology of pyrosequencing and gene polymorphism, applied in the field of molecular biology, can solve the problems of increased risk of lung cancer and achieve low cost, easy operation, and small sample volume

Inactive Publication Date: 2013-01-30
CENT SOUTH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

TERT wild-type homozygotes have a significantly higher risk of developing lung cancer than mutated genotypes

Method used

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  • TERT gene polymorphism detection kit through pyrosequencing method, and method thereof
  • TERT gene polymorphism detection kit through pyrosequencing method, and method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0028] TERT -pyroF (upstream primer): 5'-TCTATCTCAGGCATCTTGACACC-3' (SEQ ID NO.2);

[0029] TERT -pyroR (downstream primer): 5'-GGGGAACAAAGGAGGAAAAG-3' (SEQ ID NO.3);

[0030] Sequencing primer: 5'-GCGGGGGCAAAGCTA-3' (SEQ ID NO.4);

[0031] 1. DNA extraction

[0032] 1.1 The preparation and inspection of reagent materials before the experiment are as follows:

[0033] (1) Check the shelf life of the kit and ensure that ethanol has been added to Wash Buffer 1 and 2, and tick the corresponding marks on the bottle; (2) Isopropanol (if not available, absolute ethanol can be used instead) and 75% ethanol ; (3) 1.5mL Eppendorf tubes and various pipette tips within the validity period of autoclaving.

[0034] 1.2 Take out the EDTA anticoagulant tube containing whole blood from the refrigerator at 4°C, and mix it upside down several times;

[0035] 1.3 Make a mark on the 1.5mL Eppendorf tube corresponding to the unique identification of the specimen;

[0036] 1.4 Pipette 900u...

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Abstract

The present invention discloses a TERT gene polymorphism detection kit through a pyrosequencing method, and a method thereof. With the kit, TERT gene polymorphism can be detected, and specifically rs2736100 (G is more than T) single nucleotide polymorphism is detected. The kit comprises primers represented by SEQ ID NO.2-4. According to the kit, accurate, fast and high-throughput detection can be performed on the TERT gene polymorphism so as to achieve effective prediction and prevention of susceptibility risk of lung cancer.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a kit and a method for detecting TERT gene polymorphism by pyrosequencing. Background technique [0002] Lung cancer is the number one cancer killer. In my country, more than 800,000 people die from lung cancer every year. The etiology of lung cancer has not yet been fully clarified, and it may be related to genetic and environmental factors. Studies on a large sample population found that the mutation of TERT gene (rs2736100) is closely related to the genetic susceptibility to lung cancer, and the risk of developing lung cancer in non-mutated individuals is significantly higher than that in mutated individuals (J Natl Cancer Inst. 2010,102(13):959 -71; Carcinogenesis. 2009, 30(6):987–990). TERT has three genotypes, GG, GT and TT, in which GG is wild-type homozygous, GT is heterozygous, and TT is mutant homozygous. The risk of lung cancer in TERT wild-type homozyg...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 曹杉周宏灏
Owner CENT SOUTH UNIV
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