Todd Hewitt (TH) broth (Difco) was used for routine culturing of S. gordonii DL1 and isolation and culture of mutant strains. When required for mutant isolation, agar was used to solidify TH broth. Brain heart infusion (BHI) broth (Difco) was used for biofilm biomass experiments. All experiments were incubated at 37°C with 5% CO2. To construct in-frame deletions, strains were cultured at 37°C on TH agar in an anaerobic mixed gas chamber (Coy Laboratory Products). E. coli was cultured at 37°C in LB medium (Difco) or BHI. Antibiotic concentrations in TH and BHI media were used as follows: for S. gordonii, erythromycin (THerm; 5 μg/ml) and spectinomycin (THspec; 700 μg/ml), and for E. coli, LB medium was used with spectinomycin (LBspec; 75 μg/ml) or ampicillin (100 μg/ml).

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