A microfluidic system for high-speed reproducible DNA sizing and quantitation
A microfluidic system for high-speed reproducible DNA sizing and quantitation
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DOI：
10.1002/(sici)1522-2683(20000101)21:1
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发表时间：
2000-01-01
期刊：
影响因子：
2.9
通讯作者：
Ilsley, D
中科院分区：
文献类型：
--
作者：
Mueller, O;Hahnenberger, K;Ilsley, D
微加工技术被用来开发一个系统，包括一次性玻璃芯片含有蚀刻通道，试剂，包括聚合物基质和大小标准，计算机控制的仪器进行电泳分离和荧光检测的双链DNA，和软件的自动数据分析。使用不同数量和大小的DNA片段、缓冲液成分和盐浓度的样品，验证了系统性能的分离和定量再现性。已经证明了微流控系统用于DNA分析的几种应用，例如聚合酶链反应（PCR）产物、质粒引物的大小测定和通过限制性片段长度多态性（RFLP）作图检测点突变。
Microfabrication technology was used to develop a system consisting of disposable glass chips containing etched channels, reagents including polymer matrix and size standards, computer-controlled instrumentation for performing electrophoretic separations and fluorescence detection of double-stranded DNA, and software for automated data analysis. System performance was validated for separation and quantitation reproducibility using samples varying in amount and size of DNA fragments, buffer composition, and salt concentrations Several applications of the microfluidic system for DNA analysis have been demonstrated, such as of polymerase chain reaction (PCR) products, sizing of plasmid digests, and detection of point mutations by restriction fragment length polymorphism (RFLP) mapping.