Microfluidic PCR Device Using Two-stage Thermal Cycling

• Main Authors: Wen-Shian Liang, 梁文憲
• Other Authors: Chi-yang Yu
• Format: Others
• Language: en_US
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/07767888509509327458

碩士 === 大同大學 === 生物工程學系(所) === 94 === Polymerase chain reaction (PCR) is normally carried out under three specific temperatures. An integrated microfluidic PCR device was developed using microfabrication techniques. The device employed a two-stage thermal cycling. By performing annealing and elongation under the same temperature, the cycle time can be decreased and the total cycle number can be increased. The 70 mm deep flow channels were casted using polydimethylsiloxane (PDMS). A chromium layer was sputtered on Corning 1737 glass then etched and used as heaters. The glass and the polymer were bonded together to form enclosed channels. Thirty PCR cycles were manufactured in this 5.5 cm×4.0 cm device. The chip successfully amplified a 298 bp fragment from D-amino acid oxidase gene (Trigonopsis variabilis) with similar efficiency when compared to a thermal cycler. The effects of fragment length and several additives (BSA, PEG, PVP, and Triton X-100) on the device where also tested.