Development Of Static Fluorescence Quantitative Pcr With Self-sensing Thermostat Chip

Chip polymerase chain reaction (PCR) has following advantages comparing to the traditional desktop PCR:smaller heat capacity, thus high cooling and heating rate; larger specific surface area, thus high heat conduction efficiency; smaller consumption of sample and reagents. Temperature control of PCR chip is an important factor for the chip performance, final size and integration. In this paper, fluorescence quantitative PCR with self sensing thermostat chip was studied based on a static thermostat chip of indium tin oxide (ITO) glass substrate.In the first chapter, bio-chips, PCR chip in particular is reviewed. Heating, cooling and temperature measurement methods in chip PCR, principle of real-time quantitative PCR technology, its characteristics and applications are introduced in details.In the second chapter, the temperature sensing and control of the chip were characterized. It was demonstrated that with self sensing, the facrication of the chip was much simplified, and the control precision and stability were all superior to those of the thermostact chip that relied on an external temperature sensor.In the third chapter, PCR with the thermostat chip was studied. Nine parallel PCR reactions in 5μL scale were demonstrated with a chip of 25 mm×20 mm size. Real time fluorescence quantitative PCR was successfully achieved with similar detection power (105 copy number) as a commercialized desktop system. Quantitation ofλDNA added in plasma was carried out with promising result.The static thermostat chip with temperature self-sensing capability is low cost, easy to fabricate, high-throughput and highly reliabile, and would find applications in the development of portable fluorescence quantitative PCR system with disposable array PCR chips.