Disc diffusion method
The antimicrobial activity of ethanol extract of betel vine was screened using disc diffusion technique.
The agar plates were prepared by pouring 15 ml of molten nutrient agar media into sterile petriplates. The plates
were allowed to solidify and 0.1 % inoculum suspension was swabbed uniformly with sterile cotton and was
allowed to stand for 15 minutes. The different dilutions of extracts (0, 20, 40, 60 and 80 %) from initial
concentration of 1 mg/ml were loaded on 6 mm autoclaved filter paper discs. The loaded disc was placed on the
surface of medium and the compound was allowed to diffuse for 5 minutes and the plates were incubated at
37°C for 24 hrs. At the end of incubation, inhibition zones formed around the disc were measured with ruler in
millimeter. These studies were performed in triplicate.