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用于法医短串联重复序列分析的塑料微器件上DNA的超快扩增。

Ultrafast amplification of DNA on plastic microdevices for forensic short tandem repeat analysis.

作者信息

Lounsbury Jenny A, Landers James P

机构信息

Department of Chemistry, University of Virginia, 409 McCormick Road, Charlottesville, VA 22904, USA.

出版信息

J Forensic Sci. 2013 Jul;58(4):866-74. doi: 10.1111/1556-4029.12162. Epub 2013 May 21.

Abstract

The majority of microfluidic devices used as a platform for low-cost, rapid DNA analysis are glass devices; however, microchip fabrication in glass is costly and laborious, enhancing the interest in polymeric substrates, such as poly (methyl methacrylate) (PMMA), as an inexpensive alternative. Here, we report amplification in PMMA polymerase chain reaction (PCR) microchips providing full short tandem repeat profiles (16 of 16 loci) in 30-40 min, with peak height ratios and stutter percentages that meet literature threshold requirements. In addition, partial profiles (15 of 16 loci) were generated using an ultrafast PCR method in 17.1 min, representing a ~10-fold reduction in reaction time as compared to current amplification methods. Finally, a multichamber device was demonstrated to simultaneously amplify one positive, one negative, and five individual samples in 39 min. Although there were instances of loci dropout, this device represents a first step toward a microfluidic system capable of amplifying more than one sample simultaneously.

摘要

大多数用作低成本、快速DNA分析平台的微流控设备都是玻璃设备;然而,玻璃微芯片制造成本高昂且费力,这增加了人们对聚合物基底(如聚甲基丙烯酸甲酯(PMMA))作为廉价替代品的兴趣。在此,我们报告了在PMMA聚合酶链反应(PCR)微芯片中的扩增情况,该微芯片能在30 - 40分钟内提供完整的短串联重复序列图谱(16个位点中的16个),其峰高比和拖尾百分比符合文献阈值要求。此外,使用超快速PCR方法在17.1分钟内生成了部分图谱(16个位点中的15个),与当前的扩增方法相比,反应时间减少了约10倍。最后,展示了一种多腔室设备,它能在39分钟内同时扩增一个阳性样本、一个阴性样本和五个个体样本。尽管存在位点缺失的情况,但该设备代表了迈向能够同时扩增多个样本的微流控系统的第一步。

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