Growth restriction of mannose catabolic mutant and antibiotic exposure
This protocol is extracted from research article:
Slow growth determines nonheritable antibiotic resistance in Salmonella enterica
Sci Signal, Jul 30, 2019; DOI: 10.1126/scisignal.aax3938

After overnight growth in minimal medium containing tryptophan, cells were diluted 1:200 into fresh medium and allowed to grow for 2 hours. Three milliliters of culture was collected on a 0.22 μM pore-size filter (Millipore), placed in a glass filter holder assembly with a funnel, using a vacuum pump. Cells on the filter were vacuum-washed with 10 ml of deionized water (dH2O), and filters were placed on a 50-ml conical tube containing 6 ml of fresh medium containing either glucose or mannose as the sole carbon source. Cells were allowed to grow for an additional 35 min to a concentration of 2 × 107 to 20 × 107 CFU/ml before the addition of antibiotics. Cultures were subjected to a 24-hour exposure to cefotaxime or ciprofloxacin before estimation of CFU per milliliter.

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