微流控
毛细管电泳
大肠杆菌
微通道
聚合酶链反应
分子生物学
核酸
重组酶聚合酶扩增
色谱法
DNA
多重位移放大
生物
电泳
化学
基因
DNA提取
材料科学
纳米技术
生物化学
作者
Wenwu Dong,Chunxian Tao,Bo Yang,Erika Miyake,Zhenqing Li,Dawei Zhang,Yoshinori Yamaguchi
摘要
Polymerase chain reaction (PCR) is a traditional method employed for the amplification of a target gene that has played an important role in biomolecular diagnostics. However, traditional PCR is very time-consuming because of the low-temperature variation efficiency. This work proposes a continuous-flow-PCR (CF-PCR) system based on a microfluidic chip. The amplification time can be greatly reduced by running the PCR solution into a microchannel placed on heaters set at different temperatures. Moreover, as capillary electrophoresis (CE) is an ideal way to differentiate positive and false-positive PCR products, a CE system was built to achieve efficient separation of the DNA fragments. This paper describes the process of amplification of Escherichia coli (E. coli) by the CF-PCR system built in-house and the detection of the PCR products by CE. The results demonstrate that the target gene of E. coli was successfully amplified within 10 min, indicating that these two systems can be used for the rapid amplification and detection of nucleic acids.
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