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Assay kit and method for testing CYP1B1 gene polymorphism through pyrosequencing method

A technology of pyrosequencing and gene polymorphism, which is applied in the field of molecular biology, can solve the problems of increased protein expression and decreased curative effect of paclitaxel drugs, and achieves the effect of small sample size and easy operation

Inactive Publication Date: 2012-11-28
周宏灏
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CYP1B1 C4326G mutation results in increased CYP1B1 mRNA and protein expression, resulting in reduced paclitaxel efficacy

Method used

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  • Assay kit and method for testing CYP1B1 gene polymorphism through pyrosequencing method
  • Assay kit and method for testing CYP1B1 gene polymorphism through pyrosequencing method
  • Assay kit and method for testing CYP1B1 gene polymorphism through pyrosequencing method

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Experimental program
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Embodiment 1

[0028] CYP1B1-pyroF (upstream primer): 5'-GCG CTT CTC CAG CTT TGT GCC T-3' (SEQ ID NO.2);

[0029] CYP1B1-pyroR (downstream primer): 5'- AGG TCC TTG TTG ATG AGG CCA -3' (SEQ ID NO.3);

[0030] Sequencing primer: 5'- GGT CTG TGA ATC ATG ACC -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;

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Abstract

The invention discloses an assay kit and a method for testing CYP1B1 gene polymorphism through a pyrosequencing method. The assay kit is used for testing the CYP1B1 gene polymorphism, specifically rs1056836(C) G mononucleotide polymorphism. The assay kit comprises a primer, such as SEQIDN.2-4. The assay kit can be used for testing the CYP1B1 gene polymorphism in an accurate, quick and high flux manner, and thus, the individualized drug administration of substrate anti-microtubule like chemotherapeutics and the like can be realized safely, reasonably, effectively.

Description

Technical field [0001] The present invention is a molecular biology field, which involves the detection of CYP1B1 gene polymorphic kit and method. Background technique [0002] Anti -microtuba drugs have a wide anti -cancer spectrum and one of the most widely used chemotherapy drugs.Pucinol participates in the aggregation of microtubin to generate microtubuctin -related proteins, which increases the molecular weight of microtubuin sub -units to 60-76kda.After the action of paclitaxel changes, the shape of the microtuba is changed, and it does not connect to the chromosomal, which makes the cell surface protruding, and inhibits the division of wire, and the role of anti -tumor is played.Tumor cells are more sensitive in the late and M stage of G2.Commonly used drugs include paclitaxel, Doxispanne, Changchun alkali, Changchun Skinine and Changchun Risbly.The treatment of ovarian cancer and breast cancer in paclitaxel has a good effect, and has good prospects for treating prostate c...

Claims

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Application Information

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