Heiskanen M, Karhu R, Hellsten E, Peltonen L, Kallioniemi O P, Palotie A
University of Helsinki, Finland.
Biotechniques. 1994 Nov;17(5):928-9, 932-3.
Fluorescence in situ hybridization (FISH) and pulse field gel electrophoresis (PFGE) are essential techniques in physical mapping and in positional cloning. We present a technique that utilizes agarose-embedded high molecular weight DNA prepared for PFGE as a target for FISH. The agarose blocks are melted, and the DNA is extended on a poly-L-lysine-coated microscope slide. The resulting DNA fibers appear on the slide as long straight strands and are a suitable target for high resolution FISH mapping as demonstrated here with cosmid and plasmid hybridizations.
荧光原位杂交（FISH）和脉冲场凝胶电泳（PFGE）是物理图谱构建和定位克隆中的重要技术。我们提出了一种技术，该技术利用为PFGE制备的琼脂糖包埋的高分子量DNA作为FISH的靶标。将琼脂糖块融化，使DNA在聚-L-赖氨酸包被的显微镜载玻片上伸展。所得的DNA纤维在载玻片上呈现为长直链，并且如这里用黏粒和质粒杂交所证明的那样，是高分辨率FISH图谱分析的合适靶标。
