When all the G120 portions had been collected, the flasks were placed together in a boiling water bath for 30 min. The flasks contents were again vortex mixed and then cooled in ice water for 15 min.
At that point, 10 ml potassium hydroxide 7 mol l1 was added. The flasks were placed in a shaking water bath containing
ice water and shaken for 30 min. Flasks were removed one-by-one from the ice water and 1 ml of the contents was poured into tubes containing 10 ml of acetic acid (0.5 mol l1).
Amyloglucosidase solution was added to the flasks then were subsequently mixed, and placed into a 70 C water bath for 30 min followed by 10 min in a boiling water bath.
The tubes were cooled at room temperature before the addition of 40 ml water, capped, and mixed.
The tubes were then centrifuged at 1500 g for 5 min.
This process provided the total glucose (TG) portion.