title: Detection of small, single-copy genes on protein-G-banded chromosomes by electron microscopy.
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.
authors.affiliation: Département de Pathologie, Faculté de Médecine, Université de Montréal, Québec, Canada.
authors.forename: R
authors.initials: R
authors.lastname: Fetni
authors.forename: N
authors.initials: N
authors.lastname: Lemieux
authors.forename: B
authors.initials: B
authors.lastname: Malfoy
authors.forename: B
authors.initials: B
authors.lastname: Dutrillaux
authors.forename: P E
authors.initials: PE
authors.lastname: Messier
authors.forename: C L
authors.initials: CL
authors.lastname: Richer
mesh_terms: D000911:Antibodies, Monoclonal; D002871:Chromosome Banding; D002874:Chromosome Mapping / Q000379:methods*; D002882:Chromosomes, Human, Pair 13; D015342:DNA Probes; D016161:Genes, Retinoblastoma; D006801:Humans; D016253:Microscopy, Immunoelectron; D009693:Nucleic Acid Hybridization
chemical_list: D000911:Antibodies, Monoclonal; D015342:DNA Probes