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PPAR-alpha and PPAR-gamma gene polymorphism detecting chip preparing method and use

A technology of polymorphism detection and gamma gene, which is applied in the direction of biochemical equipment and methods, measurement/testing of microorganisms, etc., to achieve important social and economic benefits and strengthen the effect of early prevention

Inactive Publication Date: 2006-08-09
ZHEJIANG UNIV
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AI Technical Summary

Problems solved by technology

Kolehmainen et al. analyzed the PPARgamma2 gene Pro12Ala polymorphism in 30 cases of obesity and found that the Ala12 allele frequency was 13.3%, and there was no difference in body weight, fat thickness and fasting serum leptin level among the genotypes[2]

Method used

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Examples

Experimental program
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Embodiment 1

[0024] Example 1: Preparation of Nucleotide Gene Chip

[0025] 1) Treatment of the chip carrier: soak the glass slide in chromic acid lotion overnight, wash with water, overnight in 25% ammonia water, and wash with water. The slides were immersed in 95% ethanol solution of aminopropyltrimethoxysilane, adjusted to pH 4.5 with glacial acetic acid, ultrasonically cleaned with 95% ethanol, dried at 160°C for 4 hours, and treated with 10% glutaraldehyde to form aldehydes.

[0026]2) Synthesis of primers and probes: A set of probes were designed for the leu162val and val227ala polymorphisms of the PPAR-α gene and the pro12ala polymorphism of the PPAR-γ gene (see Table 1). Six primers were designed and synthesized for the leu162val and val227ala polymorphisms of the PPAR-α gene and the pro12ala polymorphism of the PPAR-γ gene (see Table 2). Each primer has a Cy3 fluorescent label. After synthesis, it was deprotected with concentrated ammonia water at 55°C, cut for 15 hours, and pur...

Embodiment 2

[0028] Embodiment 2: the preparation of nucleotide gene chip

[0029] 1) Treatment of the chip carrier: soak the slide in chromic acid lotion overnight, wash with water, overnight in 24% ammonia water, and wash with water. Glass slides were immersed in 95% ethanol solution of aminopropyltrimethoxysilane, adjusted to pH 4.0 with glacial acetic acid, ultrasonically cleaned with 95% ethanol, dried at 150°C for 4 hours, and treated with 9% glutaraldehyde for aldylation.

[0030] 2) Synthesis of primers and probes: A set of probes were designed for the leu162val and val227ala polymorphisms of the PPAR-α gene and the pro12ala polymorphism of the PPAR-γ gene (see Table 1). Six primers were designed and synthesized for the leu162val and val227ala polymorphisms of the PPAR-α gene and the pro12ala polymorphism of the PPAR-γ gene (see Table 2). Each primer has a Cy3 fluorescent label. After synthesis, it was deprotected with concentrated ammonia water at 50°C, cut for 13 hours, and pur...

Embodiment 3

[0032] The preparation of embodiment 3 nucleotide gene chips

[0033] 1) Treatment of the chip carrier: soak the glass slide in chromic acid lotion overnight, wash with water, overnight in 26% ammonia water, and wash with water. The glass piece was immersed in 95% ethanol solution of aminopropyltrimethoxysilane, adjusted the pH value to 4.2 with glacial acetic acid, ultrasonically cleaned with 95% ethanol, dried at 155°C for 5 hours, and treated with 11% glutaraldehyde for aldylation.

[0034] 2) Synthesis of primers and probes: A set of probes were designed for the leu162val and val227ala polymorphisms of the PPAR-α gene and the pro12ala polymorphism of the PPAR-γ gene (see Table 1). Six primers were designed and synthesized for the leu162val and val227ala polymorphisms of the PPAR-α gene and the pro12ala polymorphism of the PPAR-γ gene (see Table 2). Each primer has a Cy3 fluorescent label. After synthesis, it was deprotected with concentrated ammonia water at 60°C, cut fo...

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Abstract

This invention discloses a preparation method for the chip of testing polymorphism of PPAR-alpha and PPAR-gamma genes and its usage, in which, probes are set on the carrier. Advantage: realizing the test to the polymorphism of PPAR-alpha gene leu 162val and val 227ala and PPAR-gamma gene pro12 ala to be helpful to the selection of patients of the fatty or diabetes disease.

Description

technical field [0001] The invention relates to a preparation method and application of a PPAR-α and PPAR-γ gene polymorphism detection chip. Background technique [0002] Peroxisome proliferator-activated receptors (PPARs) were first discovered by Isseman et al. in 1990 when studying the mouse liver cDNA library. They are a class of nuclear transcription factors activated by ligands and belong to nuclear receptors. member of the body superfamily. The PPARs superfamily has three subtypes: PPARα, PPARγ, and PPARβ / δ, all of which are encoded by different genes. The expression levels of each subtype of PPARs are different in different tissues, and PPARα is mainly distributed in tissues with high metabolic activity, such as liver, kidney, heart, and muscle tissue. PPARγ is mainly expressed in adipose tissue and the immune system, and is closely related to insulin resistance. [0003] Yamakawa-Kobayashi K et al. found that the Val227Ala polymorphism of PPARα is associated with...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 厉有名陈韶华
Owner ZHEJIANG UNIV
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