Dijkwel P A, Hamlin J L
Department of Biochemistry, University of Virginia School of Medicine, Charlottesville 22908, USA.
Methods. 1997 Nov;13(3):235-45. doi: 10.1006/meth.1997.0523.
Neutral/neutral two-dimensional gel electrophoresis is a sensitive physical mapping technique that has been successfully used to unambiguously identify replication initiation sites in genomes of widely varying complexity in vivo. The technique exploits the fact that restriction fragments containing different classes of replicative intermediates (single forks, initiation bubbles, or termination structures) migrate to different and characteristic positions in agarose gels. The replication pattern of any region of interest can then be determined by sequential hybridization with appropriate radioactive probes from that region.
中性/中性二维凝胶电泳是一种灵敏的物理图谱技术，已成功用于在体内明确鉴定复杂性差异很大的基因组中的复制起始位点。该技术利用了这样一个事实，即含有不同类型复制中间体（单叉、起始泡或终止结构）的限制性片段在琼脂糖凝胶中迁移到不同的特征位置。然后，通过与来自该区域的适当放射性探针进行顺序杂交，可以确定任何感兴趣区域的复制模式。
