Biémont Christian, Monti-Dedieu Laurence, Lemeunier Françoise
Biométrie et Biologie Evolutive, Université Lyon 1, France.
Methods Mol Biol. 2004;260:21-8. doi: 10.1385/1-59259-755-6:021.
In situ hybridization is particularly appropriate for mapping specific DNA sequences on polytene chromosomes of Drosophila and other dipterans. This technique is based on the recognition and binding of one labeled sequence (the probe) to homologous sequences on chromosomes fixed on a microscope slide. The probes are labeled with biotin or other nonradioactive products, and the probe signal can be detected as a thin line on the chromosomes, following the shape of the classical Giemsa-stained chromosome bands, thus allowing the detection of TE insertions within the range of 50 to 200 kb. In our laboratory we work on many individuals from natural populations, and as a result we process high numbers of slides hybridized with various DNA probes of transposable elements every day. Therefore, the in situ hybridization technique we use is a simplification of earlier published protocols. This chapter presents our simplified standard in situ hybridization protocol for labeling polytene chromosomes of Drosophila with biotin and a fluorescence stain (FISH).
原位杂交特别适用于在果蝇和其他双翅目昆虫的多线染色体上绘制特定DNA序列。该技术基于一个标记序列（探针）与固定在显微镜载玻片上的染色体上的同源序列的识别和结合。探针用生物素或其他非放射性产物标记，并且可以按照经典吉姆萨染色染色体带的形状，将探针信号检测为染色体上的一条细线，从而能够检测50至200 kb范围内的转座子插入。在我们实验室，我们研究来自自然种群的许多个体，因此我们每天要处理大量与各种转座元件DNA探针杂交的载玻片。因此，我们使用的原位杂交技术是对早期发表的方案的简化。本章介绍了我们用生物素和荧光染料（荧光原位杂交）标记果蝇多线染色体的简化标准原位杂交方案。
