Abstract: Methanol extract of the root of Cochlospermum tinctorium was evaluated for antibacterial activities using hole-in-plate bioassay technique against Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Corynbacterium ulcerans, Proteus mirabilis and Shigella dysentriae using ciprofloxacin (10 μg mL-1) and gentamicin (10 μg mL-1) as reference standards. The extract was active on all the test organisms at concentration of 2000 μg mL-1. The activity of the extract against S. dysentriae was found to be more potent with MIC 100 and MBC 500 μg mL-1. Time kill studies showed that the antibacterial activities were time dependent. Phytochemical screening revealed the presence of alkaloids, flavonoids, tannins and cardiac glycosides. These phytochemicals could be responsible for the antimicrobial activities exhibited by the extract and hence justify the ethnomedicinal uses of C. tinctorium.