Microarray-in-a-tube Systemin The Primary Application Of Genetically Modified Crops And Detecting Human Papillomavirus Genotypes

Gene microarray is a kind of high flux and strong specificity in the level of gene detection platform, which is widely used in sequencing by hybridization, gene expression differences, SNP detection, drug screening, diagnosis, etc. The sensitivity of gene microarray affected by many factors, increasing the sensitivity of Microarray-in-a-tube System is necessary for the whole testing system which is optimized. This topic is aimed at the sensitivity of the detection of improving human Papillomavirus Genotypes and genetically modified soybean to reach the sensitivity of 100 copies/?l in Microarray-in-a-tube System.This main content of the present study are as follows:(1)The optimization of detection platform and integrating with amplification and detection integration systemFirstly, High sensitivity in order to establish new testing system, the effects of hybrid effect on the whole experiment optimized by three main steps which includes the PCR amplification system, the hybridization reaction and chip preparation in order to establish a new sensitivity system. Using general primer amplification of different HPV genotypes have different efficiency, PCR amplification system adds the specific amplification primer to obtain high efficiency. About PCR amplification system, its optimization have asymmetric primer design, PCR amplification research. The optimization design of the primer obtains the best specific amplification products to improve the efficiency of specific hybridization. Asymmetric PCR amplification study obtain more fluorescent tags single target sequence, which increases the amount of the sample of hybridization reaction to do a good ready for hybridization reaction. Hybrid time, hybrid material, quality control probe concentration has been optimized in order to the best effect of hybrid reaction, it adds the position of quality control, quality control to ensure the quality of gene microarray.(2)Microarray-in-a-tube System for detection platform, detection genetically modified soybeanGenetically modified soybean can be succeed with amplification gene MON89788, GTS-40-3-2, A2704-12, A5547-127 and internal standard gene Lectin in 100 copies.The experiment uses the platform of Microarray-in-a-tube System to detect the four genes, can The result shows successfully with detecting the four genes of MON89788, GTS-40-3-2, A2704-12, A5547-127and internal standard gene Lectin in 100 copies/?l,The signal-to-noise ratio of the transgenic soybean is up to the expected standard requirements.(3)Microarray-in-a-tube System for detection platform, detection HPV genotypesThis paper use the platform under 100 copies, which succeeds with PCR amplification of HPV16,18,31,33,35,39,45,51,52,56,58,59,68,73,26,53,68,6,11,40,42,43,44,61, 70,72,81,83 and detection HPV genotypes. About hybridization signal of HPV16,18,31,33, 35,39,40,45,44,61,70,72,81 of noise ratio evaluation analysis, All HPV genotypes of are approximately equal to 1.5, HPV45 of SNR is more lower, HPV hybridization signal of SNR values achieved the desired signal-to-noise ratio requirements.