Li Yuyuan, Xing Da, Zhang Chunsun
MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, South China Normal University, No. 55 Zhongshan Avenue West, Tianhe District, Guangzhou 510631, People's Republic of China.
Anal Biochem. 2009 Feb 1;385(1):42-9. doi: 10.1016/j.ab.2008.10.028. Epub 2008 Oct 26.
The ability to perform DNA amplification on a microfluidic device is very appealing. In this study, a compact continuous-flow polymerase chain reaction (PCR) microfluidics was developed for rapid analysis of genetically modified organisms (GMOs) in genetically modified soybeans. The device consists of three pieces of copper and a transparent polytetrafluoroethylene capillary tube embedded in the spiral channel fabricated on the copper. On this device, the P35S and Tnos sequences were successfully amplified within 9min, and the limit of detection of the DNA sample was estimated to be 0.005 ng microl(-1). Furthermore, a duplex continuous-flow PCR was also reported for the detection of the P35S and Tnos sequences in GMOs simultaneously. This method was coupled with the intercalating dye SYBR Green I and the melting curve analysis of the amplified products. Using this method, temperature differences were identified by the specific melting temperature values of two sequences, and the limit of detection of the DNA sample was assessed to be 0.01 ng microl(-1). Therefore, our results demonstrated that the continuous-flow PCR assay could discriminate the GMOs in a cost-saving and less time-consuming way.
在微流控装置上进行DNA扩增的能力非常有吸引力。在本研究中，开发了一种紧凑型连续流动聚合酶链反应（PCR）微流控技术，用于快速分析转基因大豆中的转基因生物（GMO）。该装置由三块铜和一根透明的聚四氟乙烯毛细管组成，毛细管嵌入在铜上制造的螺旋通道中。在该装置上，P35S和Tnos序列在9分钟内成功扩增，DNA样品的检测限估计为0.005 ng μl-1。此外，还报道了一种用于同时检测转基因生物中P35S和Tnos序列的双链连续流动PCR。该方法与嵌入染料SYBR Green I以及扩增产物的熔解曲线分析相结合。使用该方法，通过两个序列的特定熔解温度值识别温度差异，DNA样品的检测限评估为0.01 ng μl-1。因此，我们的结果表明，连续流动PCR检测可以以节省成本和耗时较少的方式区分转基因生物。
