DNA amplification fingerprinting (DAF) was used to evaluate the genetic relationships among 11 cultivars of poinsettia (Euphorbia pulcherrima Willd.). Amplification was with 10 octamer oligonucleotide primers that generated 336 DNA bands. Thirty-one percent of the bands were polymorphic and distinguished among cultivars. Genetic relationships were evaluated by cluster analysis, and the resulting dendrogram closely agreed with published cultivar relationships. Arbitrary signatures from amplification profiles (ASAP) were further used to characterize two cultivars, `Nutcracker Red' and `Peterstar Red', that were previously found to be genetically and morphologically similar, as well as five cultivars in the “Freedom” series. The DAF products generated with arbitrary octamer primers were reamplified with mini-hairpin decamer primers in these experiments. The ASAP profiles were complex and yielded a total of 231 bands, 38% of which were polymorphic and capable of distinguishing each Freedom cultivar. Five of the eight primer combinations distinguished `Nutcracker Red' from `Peterstar Red'. Thus, closely related cultivars of poinsettia can be separated using new and improved molecular fingerprinting protocols.