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连续流动式聚合酶链式反应微流控装置的实验研究
Experimental Study of Continuous-flow Polymerase Chain Reaction Microfluidics Based on Polytetrafluoroethylene Capillary
【摘要】 连续流动式聚合酶链式反应(PCR)微流控装置是以低热容量的软薄膜加热器为基础构建3个恒温带,以聚四氟乙烯毛细管微通道为PCR反应体系组建而成。报道了在毛细管微通道中小牛血清蛋白(BSA)动力学表面钝化对PCR扩增的影响,讨论了PCR混合物在毛细管微通道中的流动速度对PCR扩增的影响,在15 m in甚至10 m in内成功扩增了长度为249 bp的人类β-肌动蛋白片断,扩增时间仅10~15 m in,而传统PCR仪则需1~2 h。
【Abstract】 The polytetrafluoroethylene(PTFE) capillary-based continuous-flow polymerase chain reaction(PCR) microfluidics was constructed by using the flexible thin film heaters to form three constant temperature zones necessary for a PCR amplification,in which the PCR(mixture) could flow through the capillary microchannel continuously,thus leading to the successful completion of PCR exponential amplification.The effects of various concentrations of bovine serum albumin(BSA) in a 25 μL PCR(mixture) and flow rates of PCR(mixture) in the capillary microchannel,on continuous-flow PCR amplification,had been discussed in detail,where a PCR amplification of 249 base pair(bp) human β-actin gene fragments was successfully performed within 15 and even 10 minutes,which is several times faster than the reaction time required(1-2 h) with the conventional PCR machine.
【Key words】 Capillary; continuous-flow polymerase chain reaction; microfluidics;
- 【文献出处】 分析化学 ,Chinese Journal of Analytical Chemistry , 编辑部邮箱 ,2006年08期
- 【分类号】TQ315
- 【被引频次】4
- 【下载频次】227