ISOLATION OF OSTEOSARCOMA-ASSOCIATED AMPLIFIED DNA-SEQUENCES USING REPRESENTATIONAL DIFFERENCE ANALYSIS

Comparative genomic hybridization analysis of a primary osteosarcoma a nd its metastasis revealed two regions of DNA amplification, one at 17 p11.2-12 and one at 19q12-13. Subsequent representational difference a nalysis of the primary tumor resulted in the isolation of two distinct tumor-amplified DNA fragments originating from chromosome 19. A YAC c lone corresponding to one of the two isolated DNA fragments was used f or fluorescence in situ hybridization on normal human lymphocyte metap hases and tumor-derived nuclei. This resulted in the localization of t his YAC to 19q12-13.1 and confirmed the amplification status of the is olated fragment in the tumors, The availability of such RDA-isolated s equences may be instrumental in the seal-ch for genes relevant for tum or development. (C) 1997 1997 Wiley-Liss, Inc.