Direct detection of repetitive, whole chromosome paint and telomere DNA probes by immunogold electron microscopy.
Biotinylated repetitive, whole chromosome paint and telomere DNA probes were investigated at the electron microscope level after non-isotopic in situ hybridization and direct immunogold detection. The protocol described allowed the visualization of a biotinylated chromosome 1 specific satellite DNA probe in the light microscope without silver intensification. This sensitive method was exploited to analyse factors contributing to signal strength in immunogold chromosome painting. Furthermore, it allowed us to investigate the distribution of (TTAGGG)n telomere repeats in human metaphase chromosomes and interphase nuclei. Telomeric and internal (TTAGGG)n repeats were detected at high spatial resolution in human metaphase chromosomes. In the periphery of lymphocyte interphase nuclei, two types of telomere hybridization signals were observed. They differed remarkably in compactness, indicating two types of chromatin conformation present at the interphase telomeres in situ.
Steinmüller
J
J
Institut für Anthropologie, Universität Mainz, Germany.
Schleiermacher
E
E
Scherthan
H
H
D001483:Base Sequence; D002877:Chromosomes, Human / Q000648:ultrastructure*; D015342:DNA Probes; D006801:Humans; D017403:In Situ Hybridization; D007399:Interphase; D008214:Lymphocytes / Q000166:cytology / Q000648:ultrastructure*; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D012091:Repetitive Sequences, Nucleic Acid; D016615:Telomere / Q000648:ultrastructure
D015342:DNA Probes