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Human SLCO1B1 and ApoE (apolipoprotein E) gene polymorphism detection kit

A gene polymorphism and detection kit technology, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, etc. The effect of stable typing detection

Inactive Publication Date: 2015-08-19
WUHAN YZY MEDICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to overcome the deficiencies in the prior art and provide a detection kit for SLCO1B1 and ApoE gene polymorphisms, so as to solve the problem of gene polymorphism detection when the sample size is small in the existing gene polymorphism detection technology Problems with unsatisfactory results

Method used

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  • Human SLCO1B1 and ApoE (apolipoprotein E) gene polymorphism detection kit
  • Human SLCO1B1 and ApoE (apolipoprotein E) gene polymorphism detection kit
  • Human SLCO1B1 and ApoE (apolipoprotein E) gene polymorphism detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Preparation of SLCO1B1 and ApoE gene polymorphism detection kit of the present invention comprises the following steps:

[0063] 1. Primer and probe synthesis:

[0064] Design and synthesize 4 sets of specific primers SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4; SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11 and SEQ ID NO:12, and in SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17 and The 5' end of SEQ ID NO:19 is labeled with FAM fluorescent group, and the 3' end is labeled with NFQ-MGB non-luminescence quenching group, in SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18 and SEQ ID The 5' end of NO:20 is labeled with the VIC fluorescent group, and the 3' end is labeled with the NFQ-MGB non-luminescent quencher. The primers and probes were prepared into 100 μM stock solutions for storage.

[0065] 2. Prepare the internal standard system: design and synthesize a pair of internal standard primers designed for the human genome, the pr...

Embodiment 2

[0073] The SLCO1B1 and ApoE gene polymorphism detection kit prepared in Example 1 was used to detect the samples to be tested.

[0074] In this embodiment, the anticoagulated whole blood samples of 23 patients with hyperlipidemia or hyperlipidemia complications were collected, genomic DNA was extracted therefrom, and the SLCO1B1 and ApoE gene polymorphism detection kits obtained in Example 1 were used to detect the polymorphism of the samples to be tested. Gene polymorphisms of SLCO1B1 and ApoE.

[0075] 1. Genomic DNA extraction from blood samples

[0076] Take 300 μl of whole blood, add 900 μl of cell lysate CL, invert and mix well, let stand for 5 minutes, centrifuge at 10,000 rpm (11,500×g) for 1 minute, absorb the supernatant, leave the cell pellet, add to the cell pellet collected by centrifugation 200μl Buffer GS, shake until thoroughly mixed. Add 20 μl Proteinase K solution and mix well. Add 200 μl buffer GB, mix thoroughly by inversion, place at 56°C for 10 minutes...

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Abstract

The invention provides a human SLCO1B1 and ApoE (apolipoprotein E) gene polymorphism detection kit, comprising a PCR (polymerase chain reaction) buffer solution, dNTP (deoxy-ribonucleoside triphosphate), MgCl2, four groups of specific primers, four groups of specific probes, an internal standard system, HotStart Taq enzyme and UNG (uracil-N-glycosylase) enzyme. The detection kit has the advantages of high specificity, high sensitivity, ease and quickness of operation, high throughput, safety, objectiveness of result interpretation, and the like.

Description

technical field [0001] The invention relates to a human SLCO1B1 and ApoE gene polymorphism detection kit, in particular to a kit for rapidly and effectively detecting SLCO1B1 and ApoE gene polymorphisms in a small amount of samples. Background technique [0002] Statins are trihydroxytrimethylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, and have become the most widely used lipid-lowering drugs in the world since they came out. Vascular disease also played a significant role. Statins reduce the biosynthesis of cholesterol by competitively inhibiting HMG-CoA (trihydroxytrimethylglutaryl coenzyme A) reductase, block intracellular valonate metabolic pathway, and reduce intracellular cholesterol synthesis, thereby Feedback stimulates the number and activity of low-density lipoprotein receptors on the surface of cell membranes (mainly liver cells), increases the clearance of serum cholesterol, reduces the level, and then achieves the effect of lowering blood lipids. Drug ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 张喆叶婷张浩周鹏飞
Owner WUHAN YZY MEDICAL SCI & TECH
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