A proprietary multiple-DNA synthesis instrument is employed in Phase I of this project to synthesize the library of 3,314 nonamer oligonucleotides as specified for use in high throughput genome sequencing by a continuous "primer walkine' approach. The oligonucleotides are synthesized at a rate of 100 per day, affinity-purified by RPI reverse-phase cartridge chromatography, quantified and aliquoted into a "stock library" (approximately 99% of the material) and a "test library" (approximately 1% of each nonamer, or 1.5 pg/tube, sufficient for 30 sequencing reactions per primer). The test library is shipped to the Upjohn Company for testing of a proposed high throughput sequencing strategy.Anticipated Results/Potential Commercial Applications as described by the awardee:If this project is carried through to Phases II and III, a unique and valuable capability to synthesize, manage, and distribute oligonucleotide libraries for genome sequencing and mapping will have been developed. Oligonucleotide libraries should enable higher throughput and lower cost acquisition of genomic sequence information.
