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HLA-DQB1 gene typing DNA micro-array chip reagent kit

A technology of HLA-DQB1 and microarray chip, which is applied in the fields of genetic engineering, plant gene improvement, microbial measurement/inspection, etc. It can solve problems such as primer detection error, specificity reduction, and low detection throughput.

Active Publication Date: 2009-07-22
GUANGZHOU DARUI BIOTECH
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

At the same time, these methods also have some technical problems, such as detection errors caused by too many primers, specificity reduction caused by single temperature of probe hybridization, low detection throughput, etc.

Method used

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  • HLA-DQB1 gene typing DNA micro-array chip reagent kit
  • HLA-DQB1 gene typing DNA micro-array chip reagent kit
  • HLA-DQB1 gene typing DNA micro-array chip reagent kit

Examples

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Embodiment 1

[0022] Method 1: Design and Synthesis of HLA-DQB1 Genotyping DNA Microarray PCR Primers and Oligonucleotide Probes

[0023] In order to use specific PCR to amplify the second exon of the HLA-DQB1 locus, design specific 5' primers P1 and 3' primers P2-1, P2-1, P2-1 at both ends of the second exon , P2-1 was fluorescently labeled with CY3. See Table 2 for primer sequences.

[0024] Aiming at the polymorphism of the second exon gene sequence of HLA-DQB1, 18 types of typing probes were designed, as shown in Table 1.

[0025] Method 2: Preparation of HLA-DRQ1 Genotyping DNA Microarray

[0026] 18 types of typing probes and two kinds of control probes were spotted on the specific area of ​​the glass slide by using a gene chip automatic spotting instrument to make a DNA microarray.

[0027] (1) Spotting probe solution: dilute the probe into 5×SSC, 0.05% SDS solution, the final concentration is 30uM;

[0028] (2) sampling matrix: (such as figure 1 ) spotting the probes on the gla...

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Abstract

The invention relates to a gene detecting kit that is used in clinical detection, in particular to a DNA microarray chip detecting kit, which can realize subtype detection of human leukocyte antigen DQB1(HLA-DQB1) gene in high flux, high efficiency and high specificity.

Description

Technical field [0001] The invention relates to a gene detection kit for clinical detection purposes, in particular to a DNA microarray chip kit, which can detect human leukocyte antigen DQB1 (HLA-DQB1) gene with high throughput, high efficiency and high specificity. Typing detection. Background technique [0002] HLA (Human leukocyte antigen, human leukocyte antigen) is the main human histocompatibility system. Histocompatibility refers to the degree of acceptance by both the donor and the recipient when tissue or organ transplantation is performed between different individuals. The reaction caused by the incompatibility of donor and recipient tissue has been proved to be an immune response, which is induced by alloantigens on the cell surface. This antigen representing individual specificity is called transplantation antigen or histocompatibility antigen, which plays The main antigen is called the major histocompatibility system (MHS), and HLA (human leukocyte antigen) i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/12
Inventor 胡守旺张帆李明程钢何蕴韶
Owner GUANGZHOU DARUI BIOTECH
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