ITO-coated glass/polydimethylsiloxane continuous-flow PCR chip

We propose a continuous-flow polymerase chain reaction (PCR) chip using indium-tin-oxide (ITO)-coated glass/polydimethylsiloxane (PDMS) materials for DNA amplification. The continuous-flow PCR chip enables fast thermal cycling and series amplification, which are difficult to achieve in a conventional PCR or micro-chamber PCR chip. Six heaters of ITO thin films were fabricated on glass for the thermal cycling of the flowing PCR sample. The PDMS microchannel was fabricated using a negative molding method. The width and depth of the microchannel are 250 mum and 200 mum, respectively, with a total channel length of 1340 mm. The PCR chip can perform 20 cycles of amplifications. The ratio of the channel lengths for three different temperature zones, namely denaturation, annealing, and extension, is 2:2:3, respectively. Using the fabricated continuous-flow PCR chip, two DNA plasmids (720-bp pKS-GFP and 300-bp PG-noswsi) were successfully amplified.