Prefixation treatment with ethidium bromide for high resolution banding analysis of chromosomes from cultured human...

Prefixation treatment of cultured human bone marrow cells with a DNA intercalating agent, ethidium bromide (EBr), induced a dose- and time-related elongation of chromosomes. When compared with EBr-free cultures, a 2.9-fold increase in the yield of early mitotic cells with more than 400 bands per haploid set of chromosomes was achieved by simply adding 10 micrograms/ml of EBr during the last 2 hours of culture. The proportion of early mitotic cells was equal to that obtained in methotrexate synchronized cultures. Fluorescence banding methods using base composition specific agents actinomycin D/DAPI for AT base pairs and chromomycin A3/distamycin A for GC suggested that EBr does not have base specificity, because EBr did not alter the banding patterns of chromosomes obtained with these staining procedures.