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Dynamic G- and R-banding of human chromosomes for electron microscopy

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Abstract

Synchronized human lymphocytes were exposed to 5-bromo-2′-deoxyuridine (BrdUrd) for incorporation in either G-or R-bands. The substituted bands were revealed by monoclonal anti-BrdUrd antibodies disclosed with either gold-labeled antibodies or with the protein A-gold complex. Sharp G-or R-banding, specific for electron microscopy (EM), was obtained. These banding patterns, referred to as GB-AAu (G-bands by BrdUrd using Antibodies and gold [Au]) and RB-AAu (R-bands by BrdUrd using Antibodies and gold [Au]), resemble dynamic band patterns (GBG and RBG) much more than they do morphologic band patterns (GTG and RHG). The G- and R- band patterns allow accurate chromosome identification and karyotyping. An actual karyotype of human GB-AAu-banded chromosomes at the 750 band level, photographed in the EM, is presented. The method produces excellent band separation and band contrast. Variations in band staining intensities were noted and correlated with BrdUrd enrichment. The C-band regions were positively stained after GB-AAu banding while they were negatively stained after RB-AAu banding. Telomeres appeared heterogeneous after GB-AAu banding suggesting that part of the telomeric bands might be late replicating.

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Authors and Affiliations

  1. Département d'Anatomie, Faculté de Médecine, Université de Montréal, C.P. 6128 Succ. A, H3C 3J7, Montréal, Québec, Canada

    Régen Drouin, Paul-Emil Messier & Claude-Lise Richer

  2. Département de Pathologie et Centre de Recherche Pédiatrique, Hôpital Sainte-Justine, H3T 1C5, Montréal, Québec, Canada

    Claude-Lise Richer

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  1. Régen Drouin
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  2. Paul-Emil Messier
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  3. Claude-Lise Richer
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Drouin, R., Messier, PE. & Richer, CL. Dynamic G- and R-banding of human chromosomes for electron microscopy. Chromosoma 98, 40–48 (1989). https://doi.org/10.1007/BF00293333

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  • DOI: https://doi.org/10.1007/BF00293333

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