Microfabricated Flow-Through Device for In Situ Gene Analysis

A miniaturized and self-contained in situ genetic analyzer Integrated In Situ Analyzer-Gene (IISA-Gene) was developed to conduct PCR-based targeted sequence detection using microfluidic technology. The core element of the in situ analyzer is a microfluidic device that conducts cell lysis, DNA purification, PCR, and optical detection of amplified DNA fragments. In this device, a standard three-step SYBR green PCR amplification with a pair of oligonucleotide primers is conducted using environmental microbial DNA templates eluted from glass beads packed in a microchannel. The microfluidic device is integrated with pumping components and control electronics for real-field deployments. Here, the performance of the microfluidic device and results of in situ operation of the IISA-Gene in deep-sea hydrothermal areas are presented.