Dudin G, Steegmayer E W, Vogt P, Schnitzer H, Diaz E, Howell K E, Cremer T, Cremer C
Institut für Angewandte Physik I der Universität, Heidelberg, Federal Republic of Germany.
Hum Genet. 1988 Oct;80(2):111-6. doi: 10.1007/BF00702851.
Chromosomes were isolated from Chinese hamster x human hybrid cell lines containing four and nine human chromosomes. Human genomic DNA was biotinylated by nick translation and used to label the human chromosomes by in situ hybridization in suspension. Streptavidin was covalently coupled to the surface of magnetic beads and these were incubated with the hybridized chromosomes. The human chromosomes were bound to the magnetic beads through the strong biotin-streptavidin complex and then rapidly separated from nonlabeled Chinese hamster chromosomes by a simple permanent magnet. The hybridization was visualized by additional binding of avidin-FITC (fluorescein) to the unoccupied biotinylated human DNA bound to the human chromosomes. After magnetic separation, up to 98% of the individual chromosomes attached to magnetic beads were classified as human chromosomes by fluorescence microscopy.
从含有4条和9条人类染色体的中国仓鼠x人类杂交细胞系中分离出染色体。通过缺口平移法对人类基因组DNA进行生物素化,并用于在悬浮液中通过原位杂交标记人类染色体。链霉亲和素与磁珠表面共价偶联,然后将其与杂交后的染色体一起孵育。人类染色体通过强大的生物素-链霉亲和素复合物与磁珠结合,然后通过简单的永久磁铁迅速与未标记的中国仓鼠染色体分离。通过将抗生物素蛋白-异硫氰酸荧光素(荧光素)额外结合到与人类染色体结合的未占据的生物素化人类DNA上,使杂交可视化。磁分离后,通过荧光显微镜观察,附着在磁珠上的个体染色体中高达98%被归类为人类染色体。