GENOME SEQUENCING CENTER

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The proposed Genome Sequencing Center will expand the sequencing capacity of the ongoing multiplex sequencing project at CRI. An additional focus will be the development of new technology and software in order to significantly increase throughput. Forty individuals will be involved in two projects and two cores. A summary of their goals follows. Project 1: Automated Multiplex Sequencing. The overall goal is to sequence 2.4, 4, 6.4, 11.3 and 20 Mb contiguous regions in the five consecutive years of the project. The increased throughput will be accomplished by automating both laboratory and computer procedures. The M. leprae and M. tuberculosis genomes will be completed first. The sequencing target will then shift to human chromosome 10q24-q26 and its mouse syntenic regions. Overall, 24 individuals will participate in this project. The projected cost in Year 5 is approximately 25 cents per base. Project 2: Infrared Detection and Automation. This project will focus on automated high throughput hybridization and infrared fluorescence detection. The use of infrared fluorescence will significantly increase throughput, simplify quality control, and reduce costs by eliminating the use of radioactive chemicals and films. By integrating automatic hybridization and detection, all manual processes in the hybridization/detection component of multiplex sequencing will be eliminated. Over the course of five years, six machines for automated hybridization and detection will be built, enabling raw data generation at a rate of more than 500 Mb per year. Overall, 7 individuals will participate in this project. Informatics Core. The overall, 5-year objective is to develop and implement software that is capable of sustaining a processing rate of over 1.5 Mb of raw sequence data and approximately 100 kb of finished sequence per day. The four areas to be addressed are: improving base calling routines, improving sequence assembly, automating contig editing, and developing tools to facilitate sequence analysis. Additional system administration and computing tasks include network and hardware upgrades, data backups, development of project management tools, and programming support for robotics automation. Overall, this core will involve 8 individuals.