It was predicted that in this experiment to find the environmental unknown that the genus and species of the unknown would be determined. In this semester long environmental isolate experiment the purpose was to at least identify the genus of the environmental unknown. .
Materials and Methods.
In this experiment, purification of an environmental isolate was performed to learn about the different bacteria that grow in the environment. The environmental isolate was streaked off of the bottom of a rock that was planted on the soil, using a sterile cotton swab. The environmental unknown was made pure by streaking colonies onto a trypticase soy agar (TSA) plate until all of the colonies were of uniform morphology. The first TSA plate was incubated at room temperature, 25ï‚°C, for 72 hours then refrigerated at 4ï‚°C until the next laboratory. Then a selection of a colony was taken and streaked onto one TSA plate to begin purifying the environmental unknown. The plate was then incubated at room temperature from 24-48 hours then placed in the refrigerator. Once incubated to confirm the purity a gram and acid-fast stain were preformed. The stains Crystal Violet, Gram's Iodine and Safranin were used for the gram stain. The stains Carbol Fuchsin and Methylene Blue were used for the acid fast stain. These two stains were preformed to see the configurations of the environmental unknown. This aided in the process of finding out what the genus and species of the unknown was. .
The next assigned selection of an isolated colony from the first purification plate was taken and re-streaked onto a brand new TSA plate to confirm the purification of the unknown. The plate then sat a room temperature for 24-48 hours, until the growth was then checked. Once the unknown was pure a single large colony was selected and inoculated onto two tripticase soy agar slants with a screw cap tube for storage.