IRF4 gene polymorphism detection kit through pyrosequencing method, and method thereof

A technology of pyrosequencing and gene polymorphism, which is applied in the field of molecular biology to achieve the effect of simple operation and small sample volume

Inactive Publication Date: 2013-01-30
CENT SOUTH UNIV
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Problems solved by technology

Risk factors for chronic lymphocytic l

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  • IRF4 gene polymorphism detection kit through pyrosequencing method, and method thereof

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Experimental program
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Embodiment 1

[0027] IRF4 -pyroF (upstream primer): 5'-CCGTTTTTGAGACTGATCTCG-3' (SEQ ID NO.2);

[0028] IRF4 -pyroR (downstream primer): 5'-CGGGAATTTCCATTGGAGG-3' (SEQ ID NO.3);

[0029] Sequencing primer: 5'-GGCCCTTCTCTCACG-3' (SEQ ID NO.4);

[0030] 1. DNA extraction

[0031] 1.1 The preparation and inspection of reagent materials before the experiment are as follows:

[0032] (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.

[0033] 1.2 Take out the EDTA anticoagulant tube containing whole blood from the refrigerator at 4°C, and mix it upside down several times;

[0034] 1.3 Make a mark on the 1.5mL Eppendorf tube corresponding to the unique identification of the specimen;

[0035] 1.4 Pipette 900uL Cel...

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Abstract

The present invention discloses an IRF4 gene polymorphism detection kit through a pyrosequencing method, and a method thereof. With the kit, IRF4 gene polymorphism can be detected, and specifically rs872071 (A is more than G) 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 IRF4 gene polymorphism so as to achieve effective prediction and prevention of susceptibility risk of chronic lymphocytic leukaemia.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to pyrosequencing detection IRF4 Kit and method for gene polymorphism. Background technique [0002] Chronic lymphocytic leukemia (CLL) is one of the most important cancers affecting the blood and bone marrow. In CLL, lymphocytes (a type of infection-fighting white blood cell in the body) don't work as well. Over time, defective lymphocytes accumulate and reduce the number of normal white blood cells. Risk factors for chronic lymphocytic leukemia have not been fully elucidated. Chronic lymphocytic leukemia runs in families, suggesting that genetic factors play an important role in the development of the disease. The study found that located on human chromosome 6p25.3 IRF4 rs872071 of the 3'UTR of the gene is a risk variant of chronic lymphocytic leukemia, and the rs872071 gene mutation reduces the IRF4 Expression, prevent memory B cell conversion, thereby increasing the risk o...

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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