For genetic documentation and studying the taxonomic relationships, 9 species of family Euphorbiaceae; Euphorbia peplus, E. indica, E. prostrata, E. schimperi, E. tirucalli, E. granulata, Clutia myricoides, Ricinus communis, Chrozophora oblongifolia, were investigated depending on six isozymes and different molecular markers (RAPD, SSR, ITS, ITS2, matK and rbcL). Little data was obtained through the utility of RAPD and SSR primers. The four barcoding loci did not reveal discriminatory bands for all the examined species. The three loci; ITS, matK and rbcL, discriminated E. peplus and E. tirucalli from other species within genus Euphorbia. Banding patterns of the six isozyme systems showed great variability. The UPGMA dendrogram obtained depending on the 40 isozyme bands showed different findings from that of DNA. The DNA data was more accurate than isozymes data as it grouped the three species belonging to subg. Chamaesyce (E. indica, E. prostrata and E. granulata), but isozymes was more effective in separating the subfamily Acalyphoideae.