Study On The Principle, Method And Application Of PCR Chip

Micro PCR Chip system is a newly developed research field. The micro function module, such as micro channel, micro chamber, micro pump, micro heater and sensor etc, are manufactured by the technology of Micro Electro Mechanical System (MEMS) on the substrate of glass or silicon. So PCR chip has the advantage of small size, high efficiency, low cost and small contamination contrasting with traditional PCR thermo-cycler.In this dissertation, the principle of Polymerase Chain Reaction (PCR) and development of micro PCR chip system are discussed. Then the method and application of PCR chip and detection device are studied. A new PCR chip system is also developed. PCR amplification of several DNA fragments is realized by this PCR chip and the PCR results can be in situ detected by laser induced fluorescence.The topics of this dissertation are presented below:1. The principle and characteristic of Polymerase Chain Reaction are reviewed. The classification, principle, craft and tendency of PCR chip system are discussed. On the base of it, some novel PCR chip systems are presented.2. Several different PCR chips and temperature control devices are designed including quartz glass micro fluidic PCR chip, quartz glass micro chamber PCR chip and silicon-glass micro chamber PCR chip. The chips are manufactured by MEMS technics and the craft parameters are optimized. The inner side of micro chambers is smooth, flat and straight. It is good for the experiments.3. The influence of surface treatment, polymerase amount and chip structure to the chip PCR results are studied. Rapid PCR in chip and several different DNA fragments PCR in one chip at same time are realized. The speed of chip PCR is three times of traditional method.4. Chip PCR results can be in situ detected by laser induced fluorescence. This method resolved the difficulty of minimal sample in situ detection.5. A novel integrated PCR chip system with micro heaters and micro sensors on chip is designed. The processes of photolithography, wet etching, bonding and deposition of Ti/Pt alloy for micro heaters and sensors have been finished. Now assembly, circuit design and programming for temperature control are in process.The study of this dissertation showed that PCR chip has some advantages such as small size, high speed and low cost. However, much work must be developed to realize concrete application of integrate PCR chip and assemble several lab-on-chip parts to form a micro total analysis system (u-TAS).