Jefferson Andrew, Volpi Emanuela V
Department of Cardiovascular Medicine, John Radcliffe Hospital, Oxford, UK.
Methods Mol Biol. 2010;659:409-26. doi: 10.1007/978-1-60761-789-1_32.
This chapter concentrates on the use of fluorescence in situ hybridization (FISH) for genomic investigations in the laboratory rat (Rattus norvegicus). The selection of protocols included in the chapter has been inspired by a comprehensive range of previously published molecular cytogenetic studies on this model organism, reporting examples of how FISH can be applied for diverse investigative purposes, varying from comparative gene mapping to studies of chromosome structure and genome evolution, to characterization of chromosomes aberrations as well as transgenic insertions. The protocols, which include techniques for the preparation of mitotic chromosomes and DNA fibers from short-term cell cultures, have been gathered through the years and repeatedly tested in our laboratory, and all together aim at providing sufficient experimental versatility to cover a broad range of cytogenetic and cytogenomic applications.
本章重点介绍荧光原位杂交（FISH）技术在实验大鼠（褐家鼠）基因组研究中的应用。本章所选用的实验方案灵感来源于此前一系列已发表的关于该模式生物的分子细胞遗传学研究，这些研究展示了FISH技术如何应用于多种研究目的，从比较基因定位到染色体结构和基因组进化研究，再到染色体畸变以及转基因插入的表征。这些实验方案包括从短期细胞培养物中制备有丝分裂染色体和DNA纤维的技术，是多年来收集并在我们实验室反复测试过的，其共同目的是提供足够的实验通用性，以涵盖广泛的细胞遗传学和细胞基因组学应用。
