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Keywords :
Animals; DNA Polymerase I/metabolism; Deoxyribonuclease I/metabolism; Deoxyribonucleotides/metabolism; Escherichia coli/enzymology; Genetic Techniques; Immunohistochemistry; Mice; Mice, Inbred C57BL; Microscopy, Electron; Tumor Cells, Cultured/metabolism/ultrastructure
Abstract :
[en] 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.
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