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Kit for detecting organic cation transporter 2 (OCT2) gene polymorphism by pyro-sequencing method and method

A pyrosequencing method and gene polymorphism technology, applied in the field of molecular biology, can solve the problems of increasing transport function, affecting the hypoglycemic effect, reducing and other problems, and achieving the effect of small sample size and easy operation.

Inactive Publication Date: 2013-01-16
周宏灏
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  • Application Information

AI Technical Summary

Problems solved by technology

[0004] For different ethnic groups, the polymorphic sites of OCT2 are different and there are also great racial differences in the allele frequency. Gene polymorphisms lead to increased or decreased transport function, which in turn affects the clearance rate of biguanides. hypoglycemic effect

Method used

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  • Kit for detecting organic cation transporter 2 (OCT2) gene polymorphism by pyro-sequencing method and method
  • Kit for detecting organic cation transporter 2 (OCT2) gene polymorphism by pyro-sequencing method and method
  • Kit for detecting organic cation transporter 2 (OCT2) gene polymorphism by pyro-sequencing method and method

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Experimental program
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Effect test

Embodiment 1

[0028] OCT2-pyroF (upstream primer): 5′-TCG GAGAAC AGT GGG GAT TTT T-3′ (SEQ ID NO. 2);

[0029] OCT2-pyroR (downstream primer): 5′-AAT TGG GCT CTT TGT GAA ATG-3′ (SEQ ID NO. 3);

[0030] Sequencing primer: 5'-TGG TTG CAG TTC ACA G-3' (SEQ ID NO.4);

[0031] 1. DNA extraction

[0032] 1.1 The preparation and inspection of reagents and 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 mark on the bottle; (2) Isopropanol (if not available, it can be replaced with absolute ethanol) 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 anticoagulation tube containing whole blood from the refrigerator at 4°C, and invert it several times to mix.

[0035] 1.3 Mark the 1.5mL Eppendorf tube corresponding to the unique identification of the specimen;

[0036] 1.4 ...

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Abstract

The invention discloses a kit for detecting organic cation transporter 2 (OCT2) gene polymorphism by a pyro-sequencing method and a method. By the kit, the OCT2 gene polymorphism, particularly rs316019(G)T) single nucleotide polymorphism is detected. The kit comprises primers which are shown as SEQ ID NO.2-4. By the kit, the accurate, quick and high-throughout detection of OCT2 gene polymorphism can be realized, so that the safe, reasonable and effective individual administration of substrates can be realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to a kit and method for detecting OCT2 gene polymorphism by pyrosequencing. Background technique [0002] Biguanide drugs commonly used are phenformin (Jiangtangling), metformin (Jiangtang tablets, Medikang, Dihuatang, Gehuazhi, etc.), succinyl, which are effective first-line drugs for diabetic patients. Its hypoglycemic mechanism is to reduce the production of glucose in the liver, promote the uptake of glucose by the muscles, and increase insulin sensitivity. [0003] Organic cation transporters (OCTs) belong to the solute transport family, mainly including OCT1, OCT2 and OCT33 isoforms. OCT2 is mostly distributed in the basolateral membrane of proximal renal tubules, and is mainly responsible for the release of commonly used clinical drugs (such as metformin, cisplatin, cimetidine, etc.), common endogenous substances (such as dopamine, norepinephrine, etc.) and some poisons tra...

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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