Research Project
6155141
The goal is to develop and implement a novel technology for high throughput, capillary based sample handling, purification, and thermocycling to achieve a ten-fold cost reduction relative to "1/4x" ABI sequencing reactions, and a production capacity of 50,000 samples per day. Purification of E. coli lysates and sequencing reactions will be accomplished by micro-volume, modular flow-through dialysis devices using commercially available ultrafiltration membranes for which "proof of principle" has already been demonstrated. The membrane capillaries will be configured in molded cassettes such that 96 or more samples can be processed in parallel, using standard micro-well plates as sample sources. During Year-1 we will develop and test the cassettes and the liquid handling stations. During Year-2, we will produce quantities of cassettes and will place systems for evaluation at GTC, the Baylor College of Medicine, and the DOE Joint Genome Institute. During Year-3 we will implement a small-format, fully automated sample handling system at the GTC Sequencing Center and will place plasmid purification modules at the other two centers. We will also collaborate with Molecular Dynamics to develop a capillary cassette loader for their new-generation microchannel sequencer. This will allow seamless integration of the entire pipeline from E. coli clones to gel separation with sub-microliter sample volumes and a minimum of plate transfers and plastic disposables.
