Deep well petri dishes

Tryptic soy broth (TSB) (Bacto TSB, BD) prepared according to manufacturer's directions was used as the culture medium for planktonic growth.
Bio lm media was prepared by adding 0.2% agar powder (AlfaAesar) to TSB media prior to sterilization. Approximately 70 ml of molten bio lm agar was poured into deep well petri dishes (Fisher) and allowed to solidify. Bacterial strain vAh ML09-119 was removed from cryogenic storage and inoculated into 25 ml TSB media and grown overnight at 30°C with shaking. A 1 ml aliquot of overnight culture was transferred to 70 ml of TSB and grown at 30°C on an orbital shaker to mid-log phase, approximately 16 hours. Bio lm agar plates were inoculated from overnight culture by stab inoculation. Plates were sealed with para lm and incubated at 30°C for 72 hours, until an adherent bacterial lm covered the agar surface. Planktonic and bio lm cultures were performed in triplicate.

Tryptic soy broth (TSB) (Bacto TSB, BD) prepared according to manufacturer's directions was used as the culture medium for planktonic growth.
Bio lm media was prepared by adding 0.2% agar powder (AlfaAesar) to TSB media prior to sterilization, and 70 ml of molten agar was poured into deep well petri dishes (Fisher), as previously described. (64).
An aliquot of vAh ML09-119 was removed from cryogenic storage and cultured in TSB overnight at 30°C on an orbital shaker. This culture was then used to prepare planktonic and bio lm cultures, as previously described (64). Brie y, 70 ml of fresh TSB media was inoculated with 1 ml of the overnight culture and grown at 30°C with shaking to mid-log phase. Bio lm agar plates were inoculated from overnight culture by stab inoculation, sealed with para lm, and incubated at 30°C for 72 hours. Planktonic and bio lm cultures were performed in triplicate.