CORE--AUTOMATION AND INSTRUMENTATION

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Description: (Adapted from the applicant's abstract) This core laboratory is devoted to the development of new automation systems for genome mapping and sequencing. Part A, automation, will develop new methods and instruments for implementation in the production sections of Projects 1 and 2, based upon currently available methodology and prototype robotics. These developments will include: contig assembly automation, a hybridization probing system, parallel oligonucleotide synthesis, developments for the Biomek, an agarose plug preparation system, injection molded plasticware, an advanced colony picking system, technology for enhanced cycle sequencing, and the GIST. The project will also investigate the manipulation, detection and analysis of nanovolume samples for future microsequencing approaches. Part B will specifically address improvements to conventional, commercially available fluorescence-based DNA sequencing, which could lead to major increases in throughput and efficiency. Specifically, this project will implement the use of improved gel formats, such as long, thin, thermally stabilized gels for increased read lengths, and wider gels to increase throughput. It is proposed to improve the data acquisition system of the automated sequencers by providing more efficient excitation and collection optics, higher gain phototubes, faster data acquisition hardware and software and ultimately, the use of a four phototube readout system, and to improve the existing base-calling software so that the sequence data generated is more useful and to develop an integrated data collection and processing system incorporating a quality control measure to monitor the performance of the sequencers in the sequencing operation of Project 2.