Research was conducted in Tissue Culture Laboratory of ICABIOGRAD in 2013. Materials used were sugarcane varieties VMC 7616 and PS 862. Callus was induced from sugarcane leaves which still curled using MS medium added with 2,4-D 3 mg/L and casein hydrolyzate (CH) 3 g/L. To increase the genetic variability of callus, cell population were mutation-induced using EMS at concentrations 0.1%, 0.3%, and 0.5% for 30, 60, and 120 minutes by immersion. Fifty callus were used in each treatment. Mutated callus was cultured back on MS medium + 2,4-D 3 mg/L + CH 3 g/L for 3 weeks to induce somaclonal variability. Sub-cultures were performed every 2 weeks until the age of 2 months. Percentage of live callus and callus colour were observed.