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. 1990;53(2-3):134-6.
doi: 10.1159/000132913.

Rapid subchromosomal localization of cosmids by nonradioactive in situ hybridization

T Kievits  1 , J G Dauwerse, J Wiegant, P Devilee, M H Breuning, C J Cornelisse, G J van Ommen, P L Pearson
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Rapid subchromosomal localization of cosmids by nonradioactive in situ hybridization

T Kievits et al. Cytogenet Cell Genet. 1990.

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.

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