Sorry for the delayed post!
What happened on Days 2 & 3 was that there was a serious yeast contamination in the cultures set up! Why this happens: Sometimes, the bacterial strain loses its ability to make the drug 9DS: The genes that make 9DS are turned down. Because 9DS fumagates the culture and keeps fungi from growing, when this happens, yeast can contaminate the media and grow.
2400: Resuspend B. subtilis picked from LB plate (Teknova) in 300 uL distilled water, let sit for ~5 minutes to denature the bacterial biofilm. Plate the solution onto a new LB plate and core 6 cores using the wide end of a 1 mL pipette tip. Into each core, deposit 100 uL of culture from respective tubes. Let grow overnight and observe. Culture 5 was very obviously contaminated with fungal growth due to pungent smell and bright yellow color, and was disposed.
Day 3: Observation of plate showed no positive cultures.
2400: Resuspend B. subtilis picked from LB plate (Teknova) in 300 uL distilled water, let sit for ~5 minutes to denature the bacterial biofilm. Plate the solution onto a new LB plate and core 5 cores using the wide end of a 1 mL pipette tip. Into each core, deposit 100 uL of culture from respective tubes. Let grow overnight and observe.
Day 4:
Observation of plate showed no positive cultures.
Up next: How to ‘reboot’ these bacteria and get them to start making 9DS again.