Preparation of mother culture

The nutrient broth medium is prepared as detailed below

Glucose : 5.0 g Peptone : 5.0 g Beef extract : 3.0 g Sodium chloride : 3.0 g Distilled water : 1000 ml

The above medium is dispensed in conical flasks and autoclaved at 15 lb pressure for 15 mts. A loop of B.subtilis is inoculated into the medium and incubated for 2 days. This serve as the mother culture.

Mass multiplication:

The nutrient broth is prepared in fermentor and sterilized at 15 lb pressure for 15 mts. Then the mother culture is added @ 1 lit / 100 lit of the medium and incubated at room temperature for 2 days. The medium containing the bacterial growth of B.subtilis is used for mixing with talc powder.