A Spatial Temperature Gradient Capillary Array Electrophoresis System For DNA Mutation Analysis With Automated Sample Introduction

Gene mutation is closely related with many diseases, such as diabetes, cancer, heart disease, etc. Temperature gradient gel electrophoresis is an ideal method for DNA mutation detection, but the analysis throughput is low. Capillary array electrophoresis has been widely used in many fields of analytical chemistry due to its high separation efficiency and analysis throughput. So we try to develop a temperature gradient capillary array electrophoresis system for DNA mutation analysis. In this paper, a simple, automated and high-throughput capillary array electrophoresis system was developed, in which a spatial temperature gradient device provided reliable temperature gradients for multi-capillary separation columns.56℃low-range control mutation standard sample was separated effectively.In Chapter1, the development of gene mutation detection techniques was reviewed, especially temperature gradient gel electrophoresis. The development of capillary array electrophoresis was also introduced. The sampling techniques and array detection techniques of capillary array electrophoresis were both summarized.In Chapter2, a spatial temperature gradient capillary array electrophoresis system with automated sample introduction for DNA mutation analysis was established. An eight-capillary array electrophoresis chip was designed and fabricated. The temperature gradient was produced for eight capillaries at the same time by a copper block heater, and the stability of the temperature gradient was investigated.14-cm Teflon AF coated silica capillaries were used as separation channels and liquid-core waveguide tubes for transmitting fluorescence signals. A two-dimensional slotted-vial array sample introduction device controlled by a personal computer was designed. The inlets of capillaries were used for sampling, and the continuous electrokinetic sample injection was achieved.56℃low-range control mutation standard sample was separated effectively within3min with only6.5nL sample consumption, and a throughput of160/h can be achieved.In Chapter3, the spatial temperature gradient capillary array electrophoresis system with automated sample introduction was summarized, and its prospective application was discussed.