Multi-chamber; Real-Time DNA anaysis in the field using microfabricated silicon chambers

摘要

During 1997, we designed and built a 10-chamber thermal cycling system which was capable of performing the polymerase chain reaction (PCR) using TaqMan~(direct R) probes. Each chamber includes a thin-film resitive heater which was deposited directly on the microfabricated silicon chamber, for efficient heating, and the design of the silcion chambers permitted efficient, forced-convective cooling with air. Each silicon chamber contained etched holes for directly exciting the TaqMan direct R probes and monitoring their fluorescence emission. OUr software, which con trolled the thermal cycling, monitored the Taqman direct R emission from each chamber of every thermal cycle, and performed automatic calling of positives for each chamber. We used this system at Dugway Proving Grounds in january, 1998, during the detectory/identifier protion of the Joint Field Trials IV. With this system, we were able to detect and identify all three organisms that were used as simulants during JFT IV; this required the analysis of 32 unknowns per day for 10 days.