High-Throughput Sequencing of Bacterial Artificial Chromosomes and Genomic DNA

60647 The shotgun DNA sequencing method is in wide use for the acquisition of draft data, but it is not followed by equally productive finishing technology. High throughput technology is needed to transform the art of finishing genome projects into a streamlined production operation. This project will develop advanced DNA sequencing reagents and protocols that are 100 to 1,000 times more efficient than current standards and apply them for the direct sequencing of Bacterial Artificial Chromosome (BAC) and genomic DNA templates. This will be accomplished by designing a model to optimize overall workflow in the high throughput environment and test it on ongoing genomic projects. Phase I demonstrated the feasibility of increasing the yield of sequencing reactions. 10,000 Fimers were synthesized and applied for direct sequencing of BAC and microbial genomic DNA. The results showed that the direct sequencing approach was suitable for further development and implementation into large scale projects. Phase II will incorporate new enzymatic and chemical tools for more efficient sequencing of microbial genomic DNA, and overall workflow will be optimized for the high throughput environment. Commercial Applications And Other Benefits as described by the awardee: Rapid, cost effective finishing of genome projects should fulfill unmet demand in academic and government projects, as well as the booming biotechnology industry. It should free genomics centers and companies from unproductive work and accelerate the overall rate of data acquisition.