In situ localization of short low- or single-copy sequences is still difficult in plants. One solution to this problem could be the use of large yeast artificial chromosomes (YACs) for fluorescence in situ hybridization. Two YACs specific for a single copy marker on the long arm of the NOR-chromosome 2 of tomato (Lycopersicon esculentum) were selected. Both probes hybridized exclusively to this chromosome, although one produced a slightly dispersed hybridization signal. Hybridization of these YACs onto potato chromosome showed a clear single locus on the homoeologous potato chromosome in both cases.