The protocol was modified from (65). Powdered, frozen cassava root (5 cm3) was mixed with 30 ml of starch extraction medium [50 mM tris-HCl (pH 8), 0.2 mM EDTA (pH 8), and 0.5% (v/v) Triton X-100] before filtering through a 100-μm nylon mesh. The filtrate was spun at 3000g for 5 min at 20°C. The supernatant was discarded, and the starch pellet was resuspended in 5 ml of extraction medium. The suspension was filtered through a 70-μm nylon mesh, and the filtrate was overlaid on a 4-ml Percoll cushion [95% (v/v) Percoll and 5% (v/v) 0.5 M tris-HCl (pH 8)] contained in a 13-ml disposable sterile tube. Samples were spun for 5 min at 2500g. The supernatant and the Percoll cushion were vacuum-aspirated, and the remaining starch pellet was resuspended in 5 ml of 0.5% (w/v) SDS dissolved in sterile water. The samples were spun for 3 min at 4500g, and the supernatant was removed. The pellet was washed twice with sterile distilled water and then washed in 80% (v/v) ethanol before desiccation under vacuum for 36 hours and dry storage at 4°C.

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