The first stage is making Nutrient Broth. Weigh out 5.8 grams of nutrient broth powder. Dissolve completely the nutrient broth powder in beaker containing 200 ml of distilled or the deionise water. Then the mixture is transfers to the vials. Those vials are autoclave and sterilise at 1 atmosphere pressure in the temperature of 121 ℃ within 15 minutes. The nutrient Broth vials are inoculate with bacteria. The 200-600 µl of bacteria are the amount of bacteria that is will inoculate into broth. These all vials then incubate for 16 to 20 …show more content…
Weigh out 16.80 grams of agar nutrient powder. Then add and dissolve agar nutrient powder in conical flask containing 600 ml of the deionise water. Both these flask are autoclave and sterilise at temperature of 121 ℃ about 20 minutes at 1 atmosphere pressure. After the autoclave is done for these flask containing the agar nutrient, it will be let cool to 50 ℃. The mixture is swirl throughly to mix agar and nutrients. Then the nutrient agar are pour and fill 2/3 full roughly 25-35 ml per petri plate and allowed to solidify for 10 to 15 minutes. Use the spreader to uniformly spread the bacteria.
The next stage is the preparation of extracts. In this experiment which using the disk diffusion technique. This experiment use Calophyllum teysmanii, Calophyllum Andersonii and Calophyllum Canum extract and their pure compounds. Each of extracts and theirs pure compunds will weigh out 0.035 grams and will be dissolve with the 96% of ethanol. The disk was soak into the extract and then placed on the nutrient agar prepared. Then all the petri dish are placed in the incubator. The diameter of the ring appears on the nutrient agar was measured to determine the antimicrobial activity of each plant extract after 16 to 20