| There are several classify standard in microarray research. According to the source, length and fabrication method of the probe, microarray can be classified oligonucleotide and cDNA microarray.Microarray is a high-through technology to gain tremendous functional information of the gene. It can use in gene expression difference research. Thousands of probes can be immobilized to the microslide by spotting machine. cDNA microarray is very popular in many research area. The principle of hybridization in microarray is similar to those of Southern and Northern blot experiment.The manufacture of Oligonucleotide microarray includes two main direction. The light-directed synthesis technology produced by Affymetrix company is a relative successful method. Another different technology, which synthesize the probe at first, then immobilize the probes to the substrate after the crosslinking, is similar to that of cDNA.The fabrication technology of cDNA and Oligonucleotide microarry include four steps: Construction of DNA array; Preparation of DNA and RNA; Hybridization of target gene and probe; Detection and analysis of the hybridization graph.Microarray still could not be widely applied in our research because of its high cost, patent protection and fabrication technology. Our pivotal work is to fabricate and optimize the two kinds of microarray technical platform with using the slide as the substrate, and to build a basic foundation in gene detection of telomerase, multidrug-resistance, and the SNP gene detection of the CYP2D6C and the CYP2C19.The main study content in section one is as following1 .The fabrication, optimization of cDNA microarray and the detection of telomerase microarray.Our aim is to research the influence factors of the immobilization. Weselected the silylated slide as substrate because of its higher immobilization rate and lower background. Many factors could influence the immobilization rate, such as terminal amino modification, the probe length, spot solution, the probe concentration, hybridization temperature, hybridization time and UV crosslinking. We optimized these influence factors respectively.We cloned the 3 -actin as the housing-keeping gene in the former study of the telomerase detection microarray. After detecting six ordinary tumor cells, we found the telomerse gene took on higher expression level in MCF-7 and HT-2 cell line. We also detected the telomerse gene of 12 human intestine cancer tissue samples, and found that 11 samples presented high expression. 2. The preliminary study of MDR cells expression chart using microarray After treating the MCF/DOX cells with LY980503, we found 7 genes presented lower expression lever in 320 genes , these genes include:V-raf, LLGL, ankyrin motif, Arg protein tyrosine kinase-binding protein, Fms-related protein tyrosine kinase 33, 90-kDa heat-shock protein gene and COX8 gene.But RTP expression increased.In the second section, our main research result is as following.1. The optimum of SNP detecting oligonucleotide microarray.The 5' end of probe modified with amino can improve the immobilization rate, though, it can not improve the hybridization capability. The space with 15t and the detected point in midterm position are the favorable conditions. We designed 15 nucleotide as the probe which can differentiate the mutation easily.2. The manufacture of the Cytochrome oxidase 2D6C(CYP2D6C) detecting microarray.Acpording to genbank information, we synthesized the mutation detection probe, it took on an obvious difference between wild and mutant type in oligo-microarray.3. The study of the ml and m2 detection oligo-microarray on the CYP2C19. After produced the SNP detecting miaroarray of ml and m2 respectively,v/e integrated the ml and m2 miaroarray which could easily detected thedifferent mutant gene kinds of the human sample.
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