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D-StrapTM DNA Sequencing Chemistry--Fidelity Systems, Inc., 7961 Cessna Avenue, Gaithersburg, MD 20879-4117; 301-527-0804
Dr. Sergei Kozyavkin, Principal Investigator
Dr. Sergei Kozyavkin, Business Official
DOE Grant No. DE-FG02-98ER82577
Amount: $750,000
A novel strategy is needed to overcome the deficiencies inherent in genomics technology while still ensuring its complete automation and high productivity. In previous work, a unique enzyme was incorporated into a sequencing reaction to enable automated reading directly from microbial genomic DNA. This technological breakthrough opens up new possibilities in genome sequencing. This project will develop reagents and protocols for a new sequencing strategy by combining these advances in automated sequencing directly from genomic DNA with available information on the evolution of genomes. In Phase I, helper proteins for direct genomic sequencing were selected, and rRNA genes were sequenced off various genomic DNA, using universal primers. Sequencing of protein-coding genes with degenerate primers was also achieved, using microbial genomic DNA as a template. During Phase II, reagents and protocols that completely sequence rRNA operons from microbial genomic DNA, will be prepared. Additionally, about 400 universal microbial proteins will be determined and primers for these particular targets will be constructed. Two thousand to four thousand common target proteins will also be selected. Primers will then be designed and tested on several prokaryotic genomes. Finally, procedures for rapid detection of hyper-variable spots in microbial genomes will be developed.
Commercial Applications and others Benefits as described by the awardee: The new reagents should impact genomics on multiple levels by eliminating unnecessary and repetitive stages from sequencing projects and encouraging new approaches to comparative genomics. The technology should also be compatible with automated operations both at sequencing factories and service facilities, as well as for individual bench researchers.