Analysis Of Agar-Well Diffusion Method

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Methodology:
The agar-well diffusion method was employed for determining the antibacterial activities using the following sequential processes:
1. Preparation of Agar containing Petri Dishes: Standard laboratory beaker was thoroughly cleaned and dried before any use. 100ml of tap water was poured into the beaker. The hot plate was switched onto number 1 and left for three minutes to prepare for heating. At the end of three minutes the beaker containing 100 ml of water was placed on hot plate. The water was allowed to heat to 100 °C which would effectively stop any bacterial growth in water. Thermometer was used to measure the temperature of water. 2.3g of Agar powder, measured by using a mass scale, was added to the water while it was being heated. The water was stirred with a rod to make it sure that the agar mixes well and forms a homogenous solution. Once the temperature of the solution rose up to 95°C, the hot plate was slowed down to 0.5 to allow the temperature to rise slowly to 100°C (Fig 3). Once the temperature reached 100°C, the beaker was moved away from the hot plate using a pair of tongs and placed onto the heatproof mat. The next step was to prepare 30 petri dishes which were provided by the lab in packed sterile form. Each petri dish was opened partially …show more content…

Filling the petri dishes with Agar solution without any consideration of bacteria exposure. Carefully open the petri dish cover however one must not expose the entire lid. It has to be improved as the exposure of the lid allows unwanted bacteria to grow in the petri dish thus creating an unfair experiment
The use of a pipette pump to insert solutions of extracts into holes. The use of a pipette pump may have been useful for larger holes, although in this case a smaller basic pipette should have been used. An unequal measurement of each solution creates an invalid experiment. There needs to be an equal amount of

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