Mercurated nucleic acid probes, a new principle for non-radioactive in situ hybridization.
This report describes the localization of specific nucleic acid sequences in interphase nuclei and metaphase chromosomes by a new hybridocytochemical method based on the use of mercurated nucleic acid probes. After the hybridization a sulfhydryl-hapten compound is reacted with the hybrids formed. A number of such ligands were synthesized and tested. A fluorescyl ligand could be used for the direct visualization of highly repetitive sequences. For indirect immunocytochemical visualization trinitrophenyl ligands were found to be more sensitive than biotinyl analogues. These ligands were applied for the detection of target sequences in metaphase chromosomes and interphase nuclei of somatic cell hybrids, human lymphoid cell lines and blood cell cultures. The sequences were in the range of high to low copy numbers. The lower limit of sensitivity is indicated by the visualization of two human unique DNA fragments (40 and 15.6 kb) in human metaphases. The method is rapid, gives consistent results and can be used for both RNA and DNA probes. Other potentials of the new principle are discussed.
D000818:Animals; D002460:Cell Line; D004247:DNA / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006241:Haptens; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology; D007399:Interphase; D008677:Metaphase; D051379:Mice; D009693:Nucleic Acid Hybridization; D009941:Organomercury Compounds
D006241:Haptens; D009941:Organomercury Compounds; D004247:DNA