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High-resolution chromosome R-banding in lymphoblastoid cell lines by the combined use of cell synchronization and ethidium bromide treatment
Authors:Mei Hua Ye  Fumiko Saito-Ohara  Tatsuro Ikeuchi
Institution:(1) Department of Cytogenetics, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, 113 Tokyo, Japan
Abstract:Summary A reliable method for obtaining high-resolution R-banded chromosomes from lymphoblastoid cell lines is described. The cell cultures are subjected to S-phase synchronization in the presence of excess thymidine (300 mgrg/ml) for 17 to 19 hr, followed by BrdU treatment (30 mgrg/ml) for 6.5 hr. Prior to harvest, they are exposed to ethidium bromide (7.5 mgrg/ml) for 1.5 hr and Colcemid (0.02 mgrg/ml) for 30 min. Using this method, high-resolution R-banded chromosomes at the 550–850 band level were obtained with frequencies at high as 70% of all mitotic cells.
Keywords:high-resolution chromosome banding  R-banding  lymphoblastoid cell lines  ethidium bromide  cell synchronization
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