Several trials were conducted with the objective of obtaining an explant for the establishment of date palm root culture in vitro. These trials included disinfecting and germinating seeds of three cultivars on several autoclaved culture media, the influence of incubation temperature on different germination parameters, and the quality of roots before excision and after culture in nutrient media. Three culture media were used: distilled water only; minimal organics that consisted of MS salts, 3% sucrose, modified White's organics, 0.01% inositol, and 0.15% activated charcoal; and 1/2 MS salts mixture, 3% sucrose, and 1/2 modified White's organics. All three media were solidified with 0.7% agar. The seeds were incubated at 25 or 35C for germination. The study revealed the difficulty of seed disinfection. We immersed seeds in 20% to 40% Clorox, with two to four drops of Tween-20, for 30 to 60 minutes and then rinsed them four to five times in deionized distilled water before culturing. The minimal organics medium supported optimal growth of excised roots, and incubation at 35C significantly improved germination. The use of 10-mm-long root tips as explants for culture initiation gave the best growth and elongation. In addition, the growth and elongation of excised root tips increased significantly as the distance from it to the apex of the cotyledonary sheath increased.