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 ≥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.