Development Of Three Novel Integrated DNA Microarrays And Detection Instruments

The microarray technique, a high-throughput detection method, has been widely applied in the fields of the gene expression research, the SNP studies and the diagnosis development, and etc. The microarray progress is a very complex process including the gene fragment amplification, gene fragment labeling, hybridization and washing, microarray signal capture, data analysis and etc. Because of its complexity and multi-step, it is quite easy to cause unstable results. So that it has a negative impact on practical application of the microarray technique. Now, an integrated instrument of polymerase chain reaction, gene chip hybridization and fluorescence detection has been developed to perform an automatic process. The instrument is able to decrease potential disturbance of operators and increase the stability of results of the SNP detection or the disease diagnosis. Again, a microarray-in-a-tube system, which can perform the microarray progress in a sealed tube, has been developed to simplify the complexity and decrease the cross contamination in microarray progress. Furthermore, in order to solve the multi-gene quantitative problem, a quantitative microarray scanner has been designed and developed based on our lab quantitative PCR microarray technique. The scanner can be used for the multi-gene quantitative detection. And a new type of quantitative microarray was designed. The detection probes were the immobilized FRET probes. The high-throughput quantitative method would increase the efficiency of gene expression research, disease research, drug development, and drug screen and disease diagnosis.1. The integrated instrument of polymerase chain reaction and microarray hybridization and fluorescence detectionTo overcome the complexity of progress of the microarray technique, an integrated microarray, which integrates polymerase chain reactions, biochip hybridization and fluorescence detection, has been designed, developed and built. The instrument is capable of executing the whole three-step process automatically. It is good for decreasing the interfering of operators. The PCR Hybridization chips have two functional areas. The polymerase chain reaction area, made of a glass capillary, has xxx tubes with fourteen-channels. The hybridization area, covered with a glass fixed onto the chip with a rubber ring, consists of fourteen chips with detectors. The linkers between them are rubber tubers, which can be pressed by the machine to hold the liquid in the glass capillary when temperature cycles. The typical running time is about 3~4 hours for an experiment with the fourteen-unit bis-function microarray.Based on the chips above, an integrated instrument of polymerase chain reaction and microarray hybridization and fluorescence detection has been designed and constructed. The integrated instrument makes all steps of the microarray be performed automatically. It consists of a precision temperature controlling system, an automatic sample injection system and a fluorescence detection system. The hardware includes a computer, an oven, a fluorescence microscope, a temperature controller, a 2-dimension stage and a washing system. The features of the system are listed as follows.