The genetic information encoded in the human genome specifies all life processes from conception to death, and contains within it the origin of over 3500 known genetic diseases. It is necessary to substantially reduce the cost and dramatically raise the speed of producing accurate DNA sequences in order to determine the sequence of the 3 billion basepairs of DNA that compose the human genome. This proposal describes a three-year project to develop, integrate and distribute into parallel sequencing projects a fluorescent sequencing instrument and software system capable of meeting these logistical and cost demands. This approach allows for the identification and removal of the major bottlenecks encountered in producing a fully automated system, without duplication of mapping, sample processing or informatics efforts ongoing at other HGP facilities. Current bottlenecks arise primarily from inefficiencies in workflow and data analysis. Specifically, we will develop and integrate into existing centers the following: 8-dye bi-directional sequencing methods; solid phase loading of an advanced electrophoresis instrument; high-throughput and low cost ready-to-run disposable microreplicated separation sub- systems; sensitive detection optics; and advanced base-calling software. These advances will enable the completion of fully automated systems including all the steps necessary to proceed from a physical map to a finished sequence with a quantifiable measure of confidence. Full process automation coupled with automated data analysis and reduced materials cost will enable a 60-fold increase in throughput with the required 10-fold decrease in cost relative to systems based on currently available technology. This project is coordinated by CuraGen Corporation and builds on the automation, data analysis, sequencing instrumentation, and fabrication experience of collaborators at Soane BioSciences, MIT. The final product of our efforts will be tested and ready for wide-spread distribution for use in sequencing the human genome.
| Simpson, J W; Ruiz-Martinez, M C; Mulhern, G T et al. (2000) A transmission imaging spectrograph and microfabricated channel system for DNA analysis. Electrophoresis 21:135-49 |
