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When the desired acidity is reached, the mixture is again sterilized and a pure yeast culture is added.
The vortexed mixture is again stored at 4 °C for 2 h for the stabilization purpose.
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The mixture was again sonicated for 2 h.
The mixture was again incubated for 30 min at 37 °C.
The mixture was again made to blend properly.
The mixture was again covered with 1.00 g of Eschka mixture.
After the assay, the mixture was again measured using the VSM.
The mixture was again evaporated to dryness and the residue dissolved with 0.5 mol L-1 HNO3.
The mixture was again degassed using the vacuum chamber and subsequently poured into a mold.
The WHCmax was adjusted to 55% once per week, and during this procedure, the soil-sludge mixture was again mixed.
Then, the mixture was again cooled to 0 °C and L-phenylalanine (1 g, 6 mmol) was added.
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