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Nucleic Acids Research, 1991, Vol. 19, No. 12 3237-3241
© 1991

GENOME STRUCTURE AND MAPPING

In situ hybridisation with fluoresceinated DNA

Joop Wiegant, Thomas Ried, Petra M. Nederlof, Mels Van der Ploeg, Hans J. Tanke and Anton K. Raap

Department of Cytochemistry and Cytometry 72 Wassenaarseweg, 2333 AL Leiden, The Netherlands

Received April 9, 1991. Revised May 8, 1991. Accepted May 8, 1991.

We have used fluorescein-11-dUTP in a nick-translation format to produce fluoresceinated human nucleic acid probes. After In situ hybridization of fluoresceinated DNAs to human metaphase chromosomes, the detection sensitivity was found to be 50 -100 kb. The feasibility and the increase In detection sensitivity of microscopic imaging of In situ hybridized, fluoresceinated DNA with an integrating solid state camera for rapid cosmld mapping is illustrated. Combination of fluoresceinated DNA with biotinated and dlgoxigenlnated DNAs allowed easy performance of triple fluorescence In situ hybridization. The potential of these techniques for DNA mapping, cytogenetics and biological doslmetry is briefly discussed.


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