The goal of the proposed project is to participate in the mouse genome sequencing effort. Toward this goal we propose to establish a high throughput genome sequencing center at the Albert Einstein College of Medicine. We will use bacterial artificial chromosome (BAC) clones from a genomic library. The genomic library was prepared at RPCI using DNA from C57B1/6J mouse strain. We have identified several hundred BAC clones corresponding to mapped expressed sequence tags. These clones or other clones designated by the consortium will be the substrates for sequencing. The goal of the sequencing center is to reach a throughput rate of one million lanes/year at the end of the proposed three year project period. Intermediate goals are to achieve a throughput rate of 250,000 lanes/year and 500,000 lanes/year at the end of years 1 and 2, respectively. In each year we propose to complete the sequence of approximately 50 percent of the BAC clones with an error rate less than 1/10,000. Sequencing will be initiated with a combination of slab gel and capillary electrophoresis and will be shifted for all capillary electrophoresis methods. To achieve these goals, the Albert Einstein College of Medicine will provide a new 5,500 sq. ft. molecular biology laboratory with the possibility of eventually expanding into a 10,000 sq. ft. laboratory. We have developed rational and feasible plans to achieve the yearly goals. These plans include methods of recruitment, training, management and quality control.
