An inexpensive microslab gel DNA electrophoresis system with real-time fluorescence detection.
An inexpensive microslab gel DNA electrophoresis system with real-time fluorescence detection.
复制标题
具有实时荧光检测功能的廉价微板凝胶 DNA 电泳系统。
DOI：
10.1002/elps.200500572
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Ugaz,VictorM
中科院分区：
文献类型：
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作者：
Chen,Xiaojia;Ugaz,VictorM
在本文中，我们描述了一种简单而强大的凝胶电泳装置的构建，该装置可用于在几乎任何实验室中进行DNA片段的尺寸选择性分离。该系统采用了一种具有新型凝胶浇铸技术的微板凝胶格式，该技术消除了对精密梳状物的需要以限定样品加载威尔斯孔。紧凑尺寸的微板凝胶格式允许使用亚微升样品体积在低电压下进行快速分离。迁移的DNA片段的真实的实时荧光检测是使用直接连接到任何具有USB接口的PC的廉价数字显微镜来完成的。通过将其白色光源替换为蓝色发光二极管（LED）并添加适当的发射滤光片，显微镜可轻松适应此应用。聚丙烯酰胺和琼脂糖凝胶均可用作分离基质。使用标准dsDNA梯表征分离性能，并且在15分钟内实现191 bp PCR产物的正确大小测定。 该系统的低成本和简单性使其非常适合在各种实验室和教育环境中使用。
In this paper, we describe the construction of a simple yet powerful gel electrophoresis apparatus that can be used to perform size‐selective separations of DNA fragments in virtually any laboratory. This system employs a microslab gel format with a novel gel casting technique that eliminates the need for delicate combs to define sample loading wells. The compact size of the microslab gel format allows rapid separations to be performed at low voltages using submicroliter sample volumes. Real time fluorescence detection of the migrating DNA fragments is accomplished using an inexpensive digital microscope that directly connects to any PC with a USB interface. The microscope is readily adaptable for this application by replacing its white light source with a blue light‐emitting diode (LED) and adding an appropriate emission filter. Both polyacrylamide and agarose gels can be used as separation matrices. Separation performance was characterized using standard dsDNA ladders, and correct sizing of a 191 bp PCR product was achieved in 15 min. The low cost and simplicity of this system makes it ideally suited for use in a variety of laboratory and educational settings.