The laboratory fruit fly Drosophila melanogaster is one of the leading models for the study of aging. Whereas several behavioral and physiological biomarkers of aging have been identified for Drosophila, lifespan remains the most robust measure of aging rate. Aging and lifespan can be modulated by genetic alterations, as well as by drugs and dietary components, to reveal basic and conserved mechanisms of aging. Here methods are presented for Drosophila lifespan assay, including media preparation, supplementation of media with various drugs, culturing of the flies, passaging flies and recording deaths, and the analysis of lifespan data.