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Abstract: Brassica sp. is the most common oil yielding plant in India. Cotyledon, root, hypocotyl, etc. collected from Brassica nigra and Brassica juncea used as explants were incubated in MS medium containing different auxins and cytokinins either singly or in combination. Explants from Brassica nigra (except hypocotyl) when incubated in media containing 0.5 mg L-1 NAA and 2.0 mg L-1 BAP or 2.0 mg L-1 Kin exhibited formation of profuse hairy root. Best response was found in 0.5 mg L-1 NAA and 2.0 mg L-1 Kin combination. Roots were cottony in appearance. 2, 4-D either singly or in combination with other cytokinins never produced hairy root. Explants incubated in medium containing 0.5 mg L-1 IBA produced very few hairy roots. Explants of Brassica juncea transferred to medium containing similar hormone combination produced green colored compact callus but without any hairy root. Total proteins extracted from both normal and hairy roots were analyzed by 12% SDS-PAGE. A high molecular weight band was observed in hairy root but not found in normal root.