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Organization chip for researching functional genome as well as preparation method and application thereof

A tissue chip and substrate technology, which is applied in the preparation of test samples, material inspection products, biological testing, etc., can solve the problem of insufficient verification and research of differentially expressed genes, the inability to verify the spatiotemporal expression of differentially expressed genes, and the inability to match the microarray technology. matching issues

Inactive Publication Date: 2008-07-30
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The ever-innovating cDNA chip and gene chip technologies are powerful means for functional genomics research. Hundreds of differentially expressed genes can be screened out through high-throughput chip technology. The verification and research of expressed genes are very insufficient
Traditional RT-PCR and fluorescent quantitative PCR can only verify the results of chip detection, but cannot verify and study the temporal and spatial expression of differentially expressed genes; traditional in situ hybridization can only be performed with one gene probe corresponding to one tissue sample Validation, cannot match high-throughput chip technology

Method used

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  • Organization chip for researching functional genome as well as preparation method and application thereof
  • Organization chip for researching functional genome as well as preparation method and application thereof
  • Organization chip for researching functional genome as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Preparation of Tissue Chip in Tissue Development Process

[0040] 1. Materials and Methods

[0041] Tissue materials are derived from the stamen tissues of wheat spikelets in the stages of pistil differentiation, connective formation, tetrad stage, uninucleate pollen stage, and binucleate pollen stage. The stamen tissues in each stage of development are repeated 25 times, and the tissue microarray The density is 100 cores / piece, and the slice thickness is 7 microns, such as figure 1 shown.

[0042] 2. Results

[0043] The loss rate of the tissue chip was less than 5%. Safranin staining showed that the microstructure of the stamen tissue was distinct, and there was no structural deformation of the tissue. The results were as follows: figure 2 shown.

Embodiment 2

[0044] Example 2 In situ hybridization of target gene and tissue chip

[0045] 1. Materials and Methods

[0046] The tissue chip was derived from the tissue chip prepared in Example 1. Analyze the gene expression profiles of stamen tissue during the stages of androgen differentiation, connective formation, tetrad stage, uninucleate pollen stage, and binucleate pollen stage, such as image 3 As shown, the target gene TCCP1 is selected, and the expression pattern of the TCCP1 gene is as follows Figure 4 shown. The TCCP1 gene was labeled with a probe, and in situ hybridization was performed with the tissue chip in Example 1.

[0047] 2. Results The results of tissue microarray in situ hybridization showed that the in situ hybridization signal was detected at the uninucleate pollen stage, and the hybridization signal was distributed in the cells of pollen grains, anther sac walls and vascular bundles; it was also detected at the three-nucleate pollen stage The hybridization s...

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Abstract

The invention relates to the biochips field, in particular to a tissue chip used for functional genome research and the preparation method as well as the application thereof. The tissue chip of the invention includes a substrate and a paraffin section arranged on the surface of the substrate, wherein, the paraffin section is provided with tissue sample points which are in different development stages and array according to the tissue dynamic development process. Because the tissue chip of the invention can be combined with a gene chip technology, the space-time expression of a difference expressive gene screened out by the chip technology in the tissue dynamic development process is validated and researched in the high-quality, economic and available way.

Description

technical field [0001] The invention relates to the field of biochips, in particular, the invention relates to a tissue chip for functional genome research, its preparation method and application. Background technique [0002] With the completion of genome sequencing of many organisms, the study of functional genomes of different organisms has developed rapidly. The ever-innovating cDNA chip and gene chip technologies are powerful means for functional genomics research. Hundreds of differentially expressed genes can be screened out through high-throughput chip technology. The verification and research of expressed genes are very insufficient. Traditional RT-PCR and fluorescent quantitative PCR can only verify the results of chip detection, but cannot verify and study the temporal and spatial expression of differentially expressed genes; traditional in situ hybridization can only be performed with one gene probe corresponding to one tissue sample Validation cannot match hig...

Claims

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Application Information

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IPC IPC(8): G01N33/48G01N1/28
Inventor 唐忠辉赵昌平张立平杨迪
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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