A high resolution study of the DNA replication patterns of Chinese hamster chromosomes using sister chromatid differential staining technique

Chinese hamster cells were grown for 1+ and 2+ cell cycles in the presence of BrdU and then treated by the sister chromatid differential staining technique (SCD). Those regions of a chromosome which had replicated twice in the presence of BrdU were pale staining and by selecting appropriate metaphase cells an accurate reconstruction of the DNA synthetic patterns was possible. A direct correlation between the staining intensity of the G bands and the order in which they replicate was found. Dark staining G bands were always the last region of a chromosome to replicate while G negative bands were first. It is concluded that each G band may be a cluster of replicons capable of initiating DNA synthesis simultaneously.