Abstract
The purpose of this study was to develop an automated in situ gene analysis device for use in extreme environmental microbiology. Bacteria or Archaea inhabiting extreme environments such as the deep-sea or the deep subsurface are major targets of gene analysis using this device. By developing a small, fully automated and integrated gene analysis device, a small ROV (Remotely Operated Vehicle) or AUV (Autonomous Underwater Vehicle) can bring the device into underwater extreme environments to analyze microbial communities without risk of contamination and time lag problems. Since a long-term analysis operation will also be available, dynamic properties of microflora can be revealed. In this study, "microfabrication techniques" originated in the field of semiconductor or micromachine engineering are applied to develop an in situ gene analysis device that is capable of flow-through PCR (polymerase chain reaction) analyses. The device is composed of a glass-based "temperature control chip", a silicone rubber-based "microchannel chip" and a pumping system. Microfabrication techniques enable the device to be miniaturized and functionally integrated. Results of evaluation experiments indicate that the developed device can amplify targeted specific DNA fragments from E. coli cells. Furthermore, flow-through PCR operations have been tried under high static pressure conditions up to 30 MPa to simulate analysis in deep-sea environments. PCR is thus possible under high pressure, and the possibility of gene analysis in deep-sea or deep subsurface environments is demonstrated.
