The objective of this research is to develop a highly automated DNA sequencing system that can be sold to molecular biology researchers interested in carrying out moderate to large scale sequencing projects. The complete system will include robotic reaction preparation, high capacity gel electrophoresis system, an automatic gel film reader and software for managing the data. The automatic gel reader is already nearing commercial introduction under a separate phase II SBIR grant. DNASTAR has already introduced sequence management software. The specific aims of this proposal concern the robotic DNA sequence reaction robot and aspects of the gel electrophoresis system. We aim for a robot to prepare 96 DNA samples in 2 to 4 hours without human intervention. As a result we expect a yield of 384 samples for gel loading in the case of radioactive sequencing or 96 if the robot is used with a fluorescence sequencing machine. Both conventional and thermal cycle sequencing protocols will be supported. The high accuracy, reproducibility and convenience afforded by the robot, combined with the other components of DNASTAR's high throughput DNA sequencing system will support the capability to determine 50,000 or more bases of raw sequence per day. We expect the robot will be able to be manufactured inexpensively enough to be placed on the market at less than $30,000.