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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Light-emitting diode induced fluorescence (LED-IF) detection design for a pen-shaped cartridge based single capillary electrophoresis system
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Light-emitting diode induced fluorescence (LED-IF) detection design for a pen-shaped cartridge based single capillary electrophoresis system

机译:基于笔形墨盒的单毛细管电泳系统的发光二极管诱导荧光(LED-IF)检测设计

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摘要

CGE is a well-established separation technique for the analysis of biologically important molecules such as nucleic acids. The inherent high resolving power, rapid analysis times, excellent detection sensitivity, and quantification capabilities makes this method favorable compared to conventional manual polyacrylamide and agarose slab gel electrophoresis techniques. In this paper we introduce a novel single-channel capillary gel electrophoresis system with LED-induced fluorescence detection also utilizing a compact pen-shaped capillary cartridge design for automatic analysis of samples from a 96-well plate. To evaluate the suitability of the system, 1000 genomic DNA(gDNA) samples were analyzed in gel filled capillaries and detected by the microball ended excitation and emission optical fiber based LED-induced fluorescence detection system. Excellent migration time reproducibility of RSD <0.75% was obtained over the course of 1000 runs. The system rapidly distinguished between intact and degraded gDNA samples, therefore provided important information if they could be used for downstream quantitative PCR processing where high-quality intact gDNA was key. We envision that this novel system design will rapidly find new applications in both research and clinical diagnostic laboratories as a highly sensitive and easy to use bio-analytical approach. ? 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
机译:CGE是一种成熟的分离技术,用于分析生物学上重要的分子(例如核酸)。与传统的手动聚丙烯酰胺和琼脂糖平板凝胶电泳技术相比,该方法固有的高解析力,快速的分析时间,出色的检测灵敏度和定量分析功能使该方法非常适用。在本文中,我们介绍了一种新颖的具有LED诱导荧光检测功能的单通道毛细管凝胶电泳系统,该系统还利用紧凑的笔形毛细管盒设计来自动分析96孔板中的样品。为了评估系统的适用性,在凝胶填充的毛细管中分析了1000个基因组DNA(gDNA)样品,并通过基于微球末端的激发和发射光纤的LED诱导的荧光检测系统进行了检测。在1000次运行过程中,获得了RSD <0.75%的出色迁移时间重现性。该系统可快速区分完整和降解的gDNA样品,因此如果它们可用于以高质量完整gDNA为关键的下游定量PCR处理,则可提供重要信息。我们设想,这种新颖的系统设计将以高度灵敏且易于使用的生物分析方法在研究和临床诊断实验室中迅速找到新的应用。 ? 2012 WILEY-VCH Verlag GmbH&Co.KGaA,魏因海姆。

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