Odoemelam Edwin C, Raghavan Nithya, Ittiprasert Wannaporn, Miller Andre, Bridger Joanna M, Knight Matty
Division of Biosciences, Brunel University, Uxbridge, Middlesex, UK.
Methods Mol Biol. 2010;659:379-88. doi: 10.1007/978-1-60761-789-1_29.
The application of fluorescence in situ hybridization (FISH) for the mapping of single copy genes onto homologous chromosome has been integral to vast number genome sequencing projects, such as that of mouse and human. The chromosomes of these organisms are well-studied and are the staple resource of most of the early studies conducted in cytogenetics. However, there are now protocols for analyzing FISH probes in a number of different organisms on both metaphase and interphase chromosomes.Here, we describe the methodologies for the chromosomal mapping of nonrepetitive (single-copy) genes of the snail Biomphalaria glabrata onto metaphase chromosomes derived from the only molluscan cell-line in existence. The technique described in this chapter was developed for the B. glabrata genome sequencing project through troubleshooting experimental procedures established for other organisms so that both the optimum resolution of metaphase chromosome and the effective hybridization of genes were achieved.
荧光原位杂交（FISH）技术在将单拷贝基因定位到同源染色体上的应用，对于大量的基因组测序项目，如小鼠和人类的基因组测序项目而言至关重要。这些生物体的染色体已得到充分研究，是细胞遗传学早期大多数研究的主要资源。然而，现在已有在多种不同生物体的中期和间期染色体上分析FISH探针的方案。在此，我们描述了将光滑双脐螺的非重复（单拷贝）基因定位到现存唯一的软体动物细胞系衍生的中期染色体上的方法。本章所述技术是为光滑双脐螺基因组测序项目开发的，通过对为其他生物体建立的实验程序进行故障排除，从而实现了中期染色体的最佳分辨率和基因的有效杂交。
