Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Preparation and use of micro-array chip for HLA-DR4 genotyping

An HLA-DR4, microarray chip technology, applied in the field of DNA microarray chips, can solve the problems of primer detection error, small detection throughput, reduced specificity and the like

Active Publication Date: 2009-01-28
DAAN GENE CO LTD
View PDF0 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Preparation and use of micro-array chip for HLA-DR4 genotyping
  • Preparation and use of micro-array chip for HLA-DR4 genotyping
  • Preparation and use of micro-array chip for HLA-DR4 genotyping

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Method 1: Design and synthesis of PCR primers and oligonucleotide probes for HLA-DR4 genotyping DNA microarray

[0022] In order to use specific PCR to amplify the second exon of the HLA-DR4 locus, design specific 5'primers P1-1, P1-2 from the end of the first intron to the beginning of the second exon. , P1-3, P1-4, P1-5 and P1-6, and design a 3'common primer P2 at the end of the second exon. The primer sequence is shown in Table 2.

[0023] According to the polymorphism of the second exon gene sequence of HLA-DR4, 17 typing probes were designed, as shown in Table 1.

[0024] Method 2: Preparation of HLA-DRB1 genotyping DNA microarray

[0025] Using a gene chip automatic spotting instrument, 17 types of typing probes and two control probes are spotted on a specific area of ​​the glass slide to make a DNA microarray.

[0026] (1) Spot probe solution: Dilute the probe into 5×SSC, 0.05% SDS solution, the final concentration is 30uM;

[0027] (2) Spot matrix: (e.g. figure 1 ) ...

Embodiment 2

[0028] Example 2: Using HLA-DR4 genotyping to detect the HLA-DR4 genotype of samples using DNA microarray

[0029] Take 10 DNA samples with known genotypes, use CY3 labeled PCR primers, the upstream and downstream primers 1:30 (molecule number ratio) asymmetric PCR method, the PCR cycle parameters are: 94℃5'; 94℃30”, 58°C 30", 72°C 30", 40 cycles; 72°C 5'.

[0030] Take a microarray slide, take 2ul each of 10 PCR products and add them to the 10 wells of the chip, then add 8ul hybridization solution (5XSSC) to each well, put the chip into the tank of the automatic hybridization washer, and set the hybridization temperature 52°C, hybridization time 40 minutes. Automatic washing and air drying after hybridization.

[0031] A GnenPix4100A scanner is used to scan the hybridization signal to obtain a scan of the hybridization result, and convert the image into data.

[0032] Using analysis software, the above data is automatically analyzed and converted into the genotype of each sample,...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a genetic testing kit used for clinical detection, in particular to the preparation of a microarray chip and a method thereof for HLA-DR4 genotyping. By adopting a DNA microarray chip technology, the kit can carry out the genotyping to HLA DR4 gene, with high pass, 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, which can type human leukocyte antigen DR4 gene with high throughput, high efficiency and high specificity. technical background [0002] HLA (Human leukocyte antigen, human leukocyte antigen) is the main human histocompatibility system. Histocompatibility refers to the degree to which both donors and recipients accept tissue or organ transplantation between different individuals. The reaction caused by the tissue incompatibility of the donor and recipient is proved to be an immune response. It is induced by the cell surface alloantigen. This individual-specific antigen is called transplantation antigen or histocompatibility antigen. The main antigen is called the major histocompatibility system (MHS), and HLA (human leukocyte antigen) is the human major histocompatibility system. The HLA gene complex is located in the 6p21.3 region of huma...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/68
Inventor 胡守旺张帆李明程钢何蕴韶
Owner DAAN GENE CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com