To inoculate the agar plates and the previously autoclaved LB broth for bacterial testing
LB broth large flask testing
- Remove the starter cultures from the shaker.
- Pipette one mL of bacteria into each of the 8 flasks containing 50 mL LB broth (already autoclaved)
- Take UV-Vis of each of the flasks
- Place the samples to be tested into the flasks of bacteria
- The films tested were PVOH alone, PVOH + glutaraldehyde, and PVOH + quaternary ammonium salt (all 4 films/percentages). All films were cut in half, so that half of the sample was used for this test and the other half was used for agar plate testing.
- One flask was left empty, to monitor the growth of bacteria without the presence of any films.
- The final flask contained the (S)-(-)-(3-Chloro-2-hydroxypropyl)trimethylammonium chloride salt. The mass of salt used was equal to half of the amount of salt used to make the highest percentage film (because the films are being cut in half) = 0.38 g. This mass was also used for agar plate testing.
- Place the flasks on a shaker at 37°C
- Every hour to hour and a half take UV-Vis measurements
Agar Plate Preparation
- 50 mL of bacteria was pipetted onto each plate
- The inoculation loop was placed in ethanol and then placed through a flame.
- The loop was then used to spread around the bacteria.
- Samples were then added to the bacteria (see above). For the samples containing half of a film, the flattest portions of the films were used. Tape was used to try and push the films flat against the surface of the plate
- The plates were then covered and placed in an oven set to 37°C overnight.
- The films will be visualized on Friday.
UV Vis Data
- Sterile techniques were used for all of these tests (flame)
- The tape was ineffective at holding the film against the plate b/c it could not stick to the plate. It also blocked portions of the sample and made visualization more difficult.