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Application of KIT signal related gene QPCR (quantitative polymerase chain reaction) chip to mouse piebaldism detection

A signal-related, mottled disease technology, applied in the biological field, can solve the problems of high cost, emphasis on high-throughput, lack of, etc., to achieve the effect of low cost, simple operation and data analysis

Inactive Publication Date: 2012-11-14
HANGZHOU NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, it has three disadvantages: 1. High cost; 2. It is difficult to detect genes expressed in low abundance; 3. It focuses on high-throughput (testing tens of thousands or even tens of thousands of genes at a time), and lacks targets such as skin and hippocampus. Expression "chips" of specific tissue types such as tissues or certain tumors
Since the basis of this software prediction is the known signal process and cannot predict the new unknown signal process, the utilization rate of the massive data of the gene chip is limited.

Method used

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  • Application of KIT signal related gene QPCR (quantitative polymerase chain reaction) chip to mouse piebaldism detection
  • Application of KIT signal related gene QPCR (quantitative polymerase chain reaction) chip to mouse piebaldism detection
  • Application of KIT signal related gene QPCR (quantitative polymerase chain reaction) chip to mouse piebaldism detection

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preparation example Construction

[0090] Preparation of 2% agarose gel: 1.0g agarose, add 50ml of 1×TAE buffer, put in a 250ml flask, heat and dissolve in a microwave oven, cool the solution to about 60°C, add Goldview TM Nucleic acid dye to a final concentration of 0.5 μg / ml, mix well and pour into the electrophoresis plate with both ends sealed with a comb. After the gel is completely solidified, remove the comb and put the gel into the electrophoresis tank. Add a sufficient amount of 1×TAE buffer just submerged to a depth of about 1 mm from the glue surface.

[0091] Add the sample and a certain amount of 6× loading buffer into the microcentrifuge tube, mix well and add the mixture to the sample tank with a micropipette, add DL2000DNAMarker to the middle lane, and electrophoresis at 100V for 40min. After electrophoresis, observe the results under ultraviolet light.

[0092] The QPCR products of some genes were electrophoresed, except that a small amount of primer dimers could be seen in the Ptpn6 gene, the...

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Abstract

The invention belongs to the biological technical field and relates to an application of a KIT signal related gene QPCR (quantitative polymerase chain reaction) chip to the mouse piebaldism detection. The application of the KIT signal related gene QPCR chip to the mouse piebaldism detection is characterized in that the chip uses genes including Akt1, Bcl2, Cbl, Ccnd3, Ccnd3-ps, Csf1, Crk, Ecm1, Fes, Fyn, Grb7, Hras1, Jak1, Kitl, Lyn, Map2k, Matk, Myb, Nr0b2, Ntrk1, Pdgfa, Pik3ca, Ppp1r13b, Prkcc, Raf1, Sh2b2, Shc1, Snai2, Soat1, Stat1 and Zhx2 as detection sites, Actb is used as the internal reference, and the QPCR chip is used for QPCR amplification in the mouse piebaldism detection. Compared with the gene chip, the KIT signal related gene QPCR chip provided by the invention has the following advantages that 1, the cost is low; 2, the related gene is designed by aiming at a certain signal path or a certain tissue, and a certain specificity is realized; and 3, the operation and the data analysis are relatively simple.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the application of a QPCR chip for detection of KIT signal-related genes in the detection of mottle disease in mice. Background technique [0002] The human KIT proto-oncogene originated from HZ4 feline sarcoma virus, located on human chromosome 4q12, in the region around the centrosome of the long arm of chromosome 4, and is an allele of the white spot dominant gene. In mice, the gene is located at about 42 cM from the centromere on chromosome 5. The full length of human KIT genomic DNA is about 89kb, including 21 exons. The protein encoded by the KIT gene is called KIT or c-kit receptor, which is a type I transmembrane glycoprotein with a molecular weight of 145kD. It belongs to the type III protein tyrosine kinase receptor superfamily member and is distributed on the cell membrane surface. Its structure is similar to macrophage colony-stimulating factor and platelet-derived growth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 吴宝金
Owner HANGZHOU NORMAL UNIVERSITY
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