In situ nick translation at the electron microscopic level: a tool for studying the location of DNAse I-sensitive regions within the cell.
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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Laboratory of Cell and Tissue Biology, University of Liège, Belgium.
D000818:Animals; D004256:DNA Polymerase I / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism*; D003854:Deoxyribonucleotides / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D005821:Genetic Techniques; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D008854:Microscopy, Electron; D014407:Tumor Cells, Cultured / Q000378:metabolism* / Q000648:ultrastructure
D003854:Deoxyribonucleotides; D004256:DNA Polymerase I; D003850:Deoxyribonuclease I