Figure EV3. Media details to generate BFG‐GI strains and pools.

Donors, recipients, and double mutants used in BFG‐GI were generated as shown in Figs 1, EV1 and EV2. This figure shows media details, optimal inoculum cell densities, and incubation times for pool‐based cultures. All incubations were at 30°C for 24 h, except for mating (12 h at 23°C) and sporulation (12 days at 21°C). Sporulation was conducted in flasks with liquid media shaking at 200 rpm. We used the following reagent concentrations: G418 = 200 μg/ml; clonNat = 100 μg/ml; canavanine = 100 μg/ml; thialysine = 100 μg/ml; hygromycin = 200 μg/ml; and 5‐FOA = 1 mg/ml. Amino acid concentrations were as described in Tong and Boone (2005).