Wildtype strains E. cloni 10G and E. coli DH10B were inoculated in 20 ml of LB medium and incubated o/n at 37°C. The cell density was then adjusted to OD600 = 1 using fresh LB medium. Subsequently, aliquots of 100 μl of the liquid culture were plated out on LB agar plates containing increasing amounts of the antibiotics TOB (Sigma), CIP (Fluka), and TMP-SMX (Sigma). After 24 h of incubation at 37°C, the agar plates were examined for grown colonies.
MIC in liquid LB medium was determined as follows. Aliquots of 5 ml of LB medium containing increasing amounts of antibiotic were inoculated with an overnight culture of the test strains to an initial OD600 of 0.01. After incubation o/n at 37°C, the OD600 was measured.
MIC in liquid LB medium was determined as follows. Aliquots of 5 ml of LB medium containing increasing amounts of antibiotic were inoculated with an overnight culture of the test strains to an initial OD600 of 0.01. After incubation o/n at 37°C, the OD600 was measured.