- To make starter culture for future use.
- Cells were inoculated from petri dish to 3mL of sterile LB in 15mL Falcon tube.
- The falcon tube was placed in incubating orbital shaker at 37°C, 230rpm, for 24 hours.
- Cell culture was centrifuged at 4°C, 4700rpm, for 30 minutes.
- Supernatent was discarded.
- Remaining pellet was stored in -80°C freezer until future use.
- With the time remaining, an 8 hour study of cell growth versus the leaking of silver particles is going to be conducted.
- The concentration of silver incorporated into the films were calculated so that if all silvers leaked out of solution, the resulting concentration of silver in media would be 100uM to 10uM. If silver leaks out of the film dramatically from day 7 to day 8, then a dramatic decrease in measured absorbance will be found.
- The leaking of silver particles into solution will be also monitored with a sample film placed in pure water or in LB. This is the controlled study. The silver conductivity of solution will be measured everyday for 8 days. The purpose of this step is to make sure that silver leaks out of the solution, and serve as a control for above experiment. Afterwards, calculations will be made to calculate the percentage of silver leaked out of the film over time.
- If silver was observed in solution from the controlled study, and no decrease in cell culture absorbance, then it indicates the amount of silver leaked into solution is not enough to inhibit bacterial cell growth.