Different extraction techniques such as Soxhlet extraction, microwave-assisted extraction, and maceration were used to prepare aqueous, ethanolic, and methanolic extracts of Pueraria tuberosa tubers separately. For a comparative study of their cytotoxic activities in-vitro, brine shrimp lethality assay was used. The ethanolic extract of P. tuberosa tubers obtained using the microwave-assisted technique exhibited the highest cytotoxicity (LC50 = 1.56 μg/mL). Vincristine sulphate (LC50 = 0.367 μg/mL) was used as the positive control. Further phytochemical analysis and in-vivo studies need to be done to determine its potential as a source of cytotoxic drug candidates.