Differential Staining Of Sister Chromatids Research Articles

A combination of sister chromatid differential staining and giemsa banding.

We report a procedure for combining sister chromatid differential staining and G banding in the same metaphase plate. Mammalian cells in culture are grown in medium containing 5-bromodeoxyuridine for two cell cycles, and conventional air-dried preparations are made. The slides are treated with a trypsin or a urea solution the same way as for regular G banding. This method is simple and fast and provides additional information for cytogeneticists.

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.

New Giemsa method for the differential staining of sister chromatids.

IF human lymphocytes1 or Chinese hamster2 cells are treated with the base analogue 5-bromodeoxyuridine (BrdU) in the latter part of the S period, Giemsa stained chromosomes exhibit a pattern of condensed and extended segments along their length. This phenomenon has been attributed to a delay in the spiralisation pattern of the late replicating regions along the chromosomes. Other experiments3 with Chinese hamster ovary (CHO) cells have shown that after two rounds of replication in the presence of BrdU or IdU, sister chromatids stain differentially with Giemsa, allowing the identification of the two chromatids, and the observation of sister chromatid exchanges (SCEs) without recourse to autoradiography. The chromatid with the bifilarly substituted DNA (BrdU substituted in both strands of DNA) is less condensed and stains more weakly with Giemsa than the unifilarly substituted sister chromatid. The yield of SCEs is approximately that observed by autoradiography.