The cultures were
then centrifuged at 300 g for 10 min, supernatant was
removed and 10 ml of 0.075 M potassium chloride
(hypotonic solution) was added to cause hypotonic
shock (during 20 min), allowing the entrance of fluids
and inducing cellular lysis. Cells were then washed and
centrifuged at 300 g for 10 min.