High-resolution studies on late-replicating segments (G + C bands) in mammalian chromosomes

High resolution of late-replicating segments (G + C bands) in chromosomes of human, cattle and water buffalo was obtained by using 5-bromodeoxyuridine incorporation early in the cell cycle, simultaneous with methotrexate treatment combined with reduced colcemid treatment and addition of ethidium bromide, which increased the proportion of prometaphase cells. Giemsa counterstaining, following fluorescence microscopy observation and treatment with 2 x SSC, improved the resolution of the banding patterns, particularly in the pericentromeric regions. Acrocentric bovine and water buffalo chromosomes, which were seen to be C-positive by fluorescence microscopy observation and C-negative after counterstaining, showed the presence of subcentromeric G-positive bands within the heterochromatic blocks of several chromosomes.