用于哺乳动物DNA的二维DNA电泳（2D-DE）
Two-Dimensional DNA Electrophoresis (2D-DE) for Mammalian DNA.
作者信息
Walter M A, Cox D W
机构信息
Research Institute, The Hospital for Sick Children, and Departments of Molecular and Medical Genetics, and Pediatrics, University of Toronto, Canada.
出版信息
Methods Mol Biol. 1992;12:285-97. doi: 10.1385/0-89603-229-9:285.
Pulsed-field gel electrophoresis (PFGE) has been successfully used to generate physical maps of regions of human chromosomes near genes or single copy probes. However, many regions of the human genome contain gene families that often include pseudogenes. Other regions contain stretches of repeats. Conventional PFGE requires a series of single copy probes, which are sometimes difficult if not impossible to find for clusters of homologous genes or repeats. If probes detect several different genes that are dispersed on a chromosome, standard PFGE techniques will not be useful for the generation of large scale restriction maps. To overcome this problem, we have developed a method of two-dimensional DNA electrophoresis (2D-DE). The 2D-DE method consists of two DNA digestion steps and two electrophoretic separation steps (shown in Fig. 1)):
脉冲场凝胶电泳（PFGE）已成功用于绘制基因或单拷贝探针附近人类染色体区域的物理图谱。然而，人类基因组的许多区域包含基因家族，其中常常包括假基因。其他区域则含有重复序列片段。传统的PFGE需要一系列单拷贝探针，对于同源基因或重复序列簇而言，有时即便并非不可能找到这些探针，也是很困难的。如果探针检测到分散在染色体上的几个不同基因，标准的PFGE技术将无助于生成大规模限制性图谱。为克服这一问题，我们开发了一种二维DNA电泳（2D-DE）方法。二维DNA电泳方法包括两个DNA消化步骤和两个电泳分离步骤（如图1所示）：
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