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In situ nick translation at the electron microscopic level: a tool for studying the location of DNAse I-sensitive regions within the cell

M Thiry

Laboratory of Cell and Tissue Biology, University of Liege, Belgium.

The in situ nick translation method was adapted to the ultrastructural level, to study the location of DNAse I-sensitive sequences within the cell. Ultra-thin sections of Lowicryl-embedded cells were incubated in a medium containing DNAse I, DNA polymerase I, and all four deoxyribonucleotides, some being biotinylated. The nick-translated sites were then visualized by an indirect immunogold labeling technique. The resulting labeling pattern is closely dependent on the DNAse I concentration in the nick-translation medium. The method reveals with great precision the specific DNAse I-sensitive regions within the nucleus. This technique can be used to discriminate between active and inactive regions of interphase chromatin.

Volume 39, Issue 6, pp. 871-874, 06/01/1991
Copyright © 1991 by The Histochemical Society


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