Agar well diffusion technique:
The 100μl of cell free culture broth was
poured into the agar well of diameter 8mm prepared in
plates containing modified Czapek Dox’s Medium. The
filtrate was allowed to diffuse into the medium for 12
hours at 4°C. The diameter of zone (mm) of Lasparaginase
activity, as indicated by the formation of
pink coloured zone around the well against the yellow
background, was measured. For further studies,
cultures showing greater enzyme production were
selected.