Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.
Wageningen University & Research, Laboratory of Genetics, Wageningen, The Netherlands.
Methods Mol Biol. 2023;2672:351-363. doi: 10.1007/978-1-0716-3226-0_22.
DNA fiber-FISH is an easy and simple light microscopic method to map unique and repeat sequences relative to each other at the molecular scale. A standard fluorescence microscope and a DNA labeling kit are sufficient to visualize DNA sequences from any tissue or organ. Despite the enormous progress of high-throughput sequencing technologies, DNA fiber-FISH remains a unique and indispensable tool to detect chromosomal rearrangements and to demonstrate differences between related species at high resolution. We discuss standard and alternative steps to easily prepare extended DNA fibers for high-resolution FISH mapping.
DNA 纤维荧光原位杂交（DNA fiber-FISH）是一种相对简单的在分子水平上对相互间的独特序列和重复序列进行作图的方法。一台标准的荧光显微镜和一个 DNA 标记试剂盒足以使任何组织或器官的 DNA 序列可视化。尽管高通量测序技术取得了巨大的进展，但 DNA 纤维荧光原位杂交仍然是一种独特且不可或缺的工具，可用于检测染色体重排，并以高分辨率展示相关物种之间的差异。我们讨论了用于制备用于高分辨率 FISH 作图的延伸 DNA 纤维的标准和替代步骤。
