Restriction Fragment Length Polymorphisms (RFLPs) were investigated in rose cultivars as a means of reliable cultivar identification. A random genomic DNA library was generated by shotgun cloning HindIII digested fragments of DNA extracted from rose cultivar Confection into pUC8 plasmid of Escherichia coli strain JM 83. Compared to genomic clones carrying low or highly repeated sequences, clones with moderately repeated sequences were most effective in cultivar identification. These clones were identified by hybridizing rose DNA fragments from the library with genomic DNA from `Confection'. Clones with moderately repeated copy sequences were used as probes to detect the presence of RFLPs by Southern hybridization of EcoRI digested genomic DNA of various rose cultivars. Several of these probes have revealed RFLPs useful in cultivar identification. By using a combination of two or more of these probes most of the rose cultivars compared at this time can be identified. A dichotomous key useful in identification of rose cultivars was prepared from RFLPs displayed by 3A9 probe.