Hybridization of radiolabeled wheat DNA probes to genomic DNA digests of compensating nullisomic-tetrasomic lines and ditelosomic lines of hexaploid wheat (Triticum aestivum L. cv. Chinese Spring) can be used to identify intergenomic RFLPs. Sixty-three PstI/BamHI genomic DNA probes and eight cDNA probes were used to determine the chromosomal locations of 223 DNA fragments that define a minimum of 189 RFLP loci. Eighty-four percent of the genomic DNA clones hybridize to fragments located in homoeologous chromosomes and 16% hybridize to fragments located in one chromosome only or to fragments located in nonhomoeologous chromosomes. All of the cDNA probes hybridize to fragments located in homoeologous chromosomes.