Kit and method for detecting CYP19A1 gene polymorphism by pyrophosphoric acid sequencing method

A technology of pyrosequencing and gene polymorphism, which is applied in the field of molecular biology, can solve the problem of loss of enzyme activity and achieve the effect of small sample size and easy operation

Inactive Publication Date: 2012-08-22
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Exemestane is a steroidal AIs with an androgen structure, which competes with androstenedione, a substrate

Method used

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  • Kit and method for detecting CYP19A1 gene polymorphism by pyrophosphoric acid sequencing method
  • Kit and method for detecting CYP19A1 gene polymorphism by pyrophosphoric acid sequencing method
  • Kit and method for detecting CYP19A1 gene polymorphism by pyrophosphoric acid sequencing method

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Embodiment 1

[0029] CYP19A1-pyroF (upstream primer): 5'-TCAAACTCTTGGCCTCTGCTTT-3' (SEQ ID NO.2);

[0030] CYP19A1-pyroR (downstream primer): 5'-TGGCCCATGGCATTTTAGG-3' (SEQ ID NO.3);

[0031] Sequencing primer: 5'-CCAAGCTAGGTGCTATT-3' (SEQ ID NO.4);

[0032] 1. DNA extraction

[0033] 11 The preparation and inspection of reagent materials before the experiment are as follows:

[0034] (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 mark on the bottle; (2) Isopropanol (if not available, it can be replaced by absolute ethanol) and 75% ethanol ; (3) 1.5mL Eppendorf tubes and various pipette tips within the validity period of autoclaving.

[0035] 1.2 Take out the EDTA anticoagulant tube containing whole blood from the refrigerator at 4°C, and mix it upside down several times;

[0036] 1.3 Make a mark on the 1.5mL Eppendorf tube corresponding to the unique identification of the specimen;

[0037] 1.4 Pipette 900u...

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Abstract

The invention discloses a kit and method for detecting CYP19A1 gene polymorphism by a pyrophosphoric acid sequencing method. The kit is used for examining the CYP19A1 gene polymorphism, particularly rs4646 (G is greater than T) single nucleotide polymorphism. The kit comprises primers shown as SEQ ID NO. 2-4. According to the kit, accurate, quick and high-flux detection of the CYP19A1 gene polymorphism can be realized, so that individual administration for realizing safe, rational and effective suppression on primer aromatase of the kit is achieved.

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 CYP19A1 gene polymorphism by pyrosequencing. Background technique [0002] Aromatase (Aromatase Enzyme) is a member of the cytochrome P450 enzyme superfamily, and the coding gene is CYP19A1, which is located at 15q211. Enzymes necessary for different stages of aldosterone, cortisol, androgen biosynthesis. CYP19A1 can catalyze the conversion of testosterone and androstenedione to estrone and estradiol, and play the final rate-limiting catalytic role in estrogen biosynthesis, so it is also called estrogen synthase. [0003] Aromatase inhibitors (Aromatase inhibitors, AIs) reduce the level of estrogen by inhibiting aromatase, thereby eliminating the stimulating effect of estrogen on tumor growth. Estrogen in postmenopausal women is mainly derived from the aromatization of androgen precursors in peripheral tissues, so this type of drug i...

Claims

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Application Information

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