Detection of small, single-copy genes on protein-G-banded chromosomes by electron microscopy. |
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Authors: | R Fetni N Lemieux B Malfoy B Dutrillaux P E Messier C L Richer |
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Affiliation: | Département de Pathologie, Faculté de Médecine, Université de Montréal, Québec, Canada. |
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Abstract: | A method for the detection by electron microscopy of chromosome banding after in situ hybridization of small, nonradioactive DNA sequences is described. Typical high-resolution G-banding is produced by adding 5-bromodeoxyuridine (BrdU) during the last part of the S-phase and by applying a monoclonal antibody against the BrdU-substituted chromosome segments, followed by the addition of protein G, but no further treatment. A protocol for in situ hybridization of small, single-copy biotinylated DNA sequences and their detection by immunogold tagging on banded chromosomes is also described. This combined approach permits high-resolution mapping of small DNA sequences and should be useful in discriminating between neighboring DNA fragments. |
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