Fluorescence in situ hybridization of YAC clones after Alu-PCR amplification.
Alu-PCR protocols were optimized for the generation of human DNA probes from yeast strains containing yeast artificial chromosomes (YACs) with human inserts between 100 and 800 kb in size. The resulting DNA probes were used in chromosome in situ suppression (CISS) hybridization experiments. Strong fluorescent signals on both chromatids indicated the localization of specific YAC clones, while two clearly distinguishable signals were observed in greater than or equal to 90% of diploid nuclei. Signal intensities were generally comparable to those observed using chromosome-specific alphoid DNA probes. This approach will facilitate the rapid mapping of YAC clones and their use in chromosome analysis at all stages of the cell cycle.
Lengauer
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Institut für Humangenetik u. Anthropologie, Universität Heidelberg, Federal Republic of Germany.
Green
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Cremer
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D001483:Base Sequence; D015825:Chromosomes, Fungal; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D015342:DNA Probes; D005453:Fluorescence; D005784:Gene Amplification; D015723:Gene Library; D015894:Genome, Human; D006801:Humans; D006822:Hybrid Cells; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid
D015342:DNA Probes; D004247:DNA