High-resolution chromosome R-banding in lymphoblastoid cell lines by the combined use of cell synchronization and ethidium bromide treatment |
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Authors: | Mei Hua Ye Fumiko Saito-Ohara Tatsuro Ikeuchi |
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Affiliation: | (1) Department of Cytogenetics, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, 113 Tokyo, Japan |
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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 g/ml) for 17 to 19 hr, followed by BrdU treatment (30 g/ml) for 6.5 hr. Prior to harvest, they are exposed to ethidium bromide (7.5 g/ml) for 1.5 hr and Colcemid (0.02 g/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. |
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Keywords: | high-resolution chromosome banding R-banding lymphoblastoid cell lines ethidium bromide cell synchronization |
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