title: Rapid subchromosomal localization of cosmids by nonradioactive in situ hybridization.
abstract: A rapid method for localizing large numbers of complete cosmids by nonradioactive in situ hybridization is described. The cosmids are nick translated in the presence of biotin-16-dUTP, incubated with an excess of sonicated human DNA, and used as a probe for in situ hybridization. Sites of hybridization are detected by successive treatments with FITC-labeled avidin and biotinylated anti-avidin antibody. Fifty-two cosmids were localized on chromosome 16 in 5 d relative to translocation breakpoints contained in two cell lines. Rapid identification of chromosome 16 was achieved by cohybridization with a chromosome 16-specific centromeric repeat probe.
authors.affiliation: Department of Human Genetics, State University of Leiden, The Netherlands.
authors.forename: T
authors.initials: T
authors.lastname: Kievits
authors.forename: J G
authors.initials: JG
authors.lastname: Dauwerse
authors.forename: J
authors.initials: J
authors.lastname: Wiegant
authors.forename: P
authors.initials: P
authors.lastname: Devilee
authors.forename: M H
authors.initials: MH
authors.lastname: Breuning
authors.forename: C J
authors.initials: CJ
authors.lastname: Cornelisse
authors.forename: G J
authors.initials: GJ
authors.lastname: van Ommen
authors.forename: P L
authors.initials: PL
authors.lastname: Pearson
mesh_terms: D002461:Cell Line, Transformed; D002874:Chromosome Mapping; D002885:Chromosomes, Human, Pair 16; D003360:Cosmids; D005260:Female; D006801:Humans; D008856:Microscopy, Fluorescence; D009693:Nucleic Acid Hybridization