A 5000. rad goat-hamster panel of 121 whole-genome radiation hybrids was generated and preliminarily characterized. A normal diploid fibroblast culture from a male Boer goat was fused with a recipient thymidine kinase-deficient hamster cell line. The generated 121 radiation hybrids were grown and produced an average of 8.4. mg of DNA per radiation hybrid. A SINE-PCR test showed that almost all radiation hybrids retained goat DNA. The retention frequencies of the 121 hybrids were preliminarily estimated using a collection of 42 unlinked molecular markers. An optimized panel of therein 90 radiation hybrids (CHRH5000) with an average retention frequency of 34.2% was screened on the basis of interspersed repetitive DNA content and chromosome retention frequency. The development of this WG-RH panel will provide a fundamental tool for advanced goat genome mapping studies and for mammalian comparative mapping. © 2012 Elsevier B.V.