Bacto tsb

Tryptic soy broth (TSB) (Bacto TSB, BD) prepared according to manufacturer’s directions was used as the culture medium for planktonic growth.
Biofilm 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 (link)].. 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 biofilm cultures, as previously described [64 (link)]. Briefly, 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. Biofilm agar plates were inoculated from overnight culture by stab inoculation, sealed with parafilm, and incubated at 30 °C for 72 h. Planktonic and biofilm cultures were performed in triplicate.

Tryptic Soy Broth (TSB) (Bacto TSB, BD) and Luria Broth (LB) prepared according to manufacturer's directions was routinely used as the culture medium for growth of vAh and Escherichia coli, respectively, with the addition of 1.5% agar powder (Alfa Aesar) for solid culture. When necessary for selection, media were supplemented with chloramphenicol (Cam) (25 μg/ml), tetracycline (Tet) (10 μg/ml), colistin (Col) (10 μg/ml), kanamycin (Kan) (30 μg/ml), or combinations of those antibiotics. Super Optimal Broth (SOB) (Difco SOB, BD) was used for culture of electrocompetent cells and Super Optimal Broth with Catabolite Repression (SOC) was used as a recovery medium following electroporation. vAh strains were cultured at 30°C and E. coli strains were cultured at 37°C.
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. An aliquot of overnight culture was transferred to 70 ml of TSB and grown at 30°C on an orbital shaker at 180 rpm to mid-log phase, ~16 h.