Rapid subchromosomal localization of cosmids by nonradioactive in situ hybridization |
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| Authors: | T Kievits J G Dauwerse J Wiegant P Devilee M H Breuning C J Cornelisse G J van Ommen P L Pearson |
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| Institution: | Department of Human Genetics, State University of Leiden, The Netherlands. |
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| 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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