54
High-Throughput
Sequencing of Bacterial Artificial Chromosomes and Genomic DNA--Fidelity Systems, Inc., 7961
Cessna Avenue, Gaithersburg, MD 20879-4117;
301-527-0804
Dr. Sergei Kozyavkin, Principal Investigator
Dr. Andrei Malykh, Business Official
DOE Grant No. DE-FG02-00ER83009
Amount:
$100,000
Wide use of the shotgun DNA
sequencing method for the acquisition of draft data is not followed by the
equally productive finishing technology. We have recently combined our proprietary biochemical and chemical
approaches and showed feasibility of rapid finishing of draft human BAC
projects with 2.3x-4.5x coverage to the high quality and contiguity standards. Development of high throughput technology
based on our methods will transform the art of finishing genome projects into a
streamlined production operation. Our
objective is the development of advanced DNA sequencing reagents and protocols
that are 100 to 1,000 times more efficient than the current standards and apply
them for the sequencing directly off BAC and genomic DNA templates. We will design a model for the optimization
of the overall workflow in the high throughput environment and test it on the
ongoing genomics projects. In Phase I
we will demonstrate feasibility of our approach to multiply the yield of
sequencing reactions. We will show how
the advanced chemistry will simplify massive sequencing of BAC and microbial
genomic DNA and make direct sequencing of larger genomes possible. We will also develop a new solution for
sample tracking and internal control.
Commercial Applications and Other Benefits as described by the
awardee: Rapid and cost effective
finishing of genome projects will fulfill unmet demand in academic and
government projects and booming biotechnology industry. It will free genomics centers and companies
from unproductive work and accelerate overall rate of data acquisition. Streamlined sequencing directly off genomic
DNA without cloning or PCR steps will find wide applications in functional and
comparative genomics.