The mid-exponential phase and growth-arrested cultures (3 mL) were generated as described above. CaCl2 was added to a final concentration of 1.25 mM, and daptomycin was added to 20 μg mL−1. The cultures were incubated at 37°C with shaking (180 rpm) for 6 h. At each time point, aliquots were taken, serially diluted 10-fold in PBS, and plated onto TSB to determine the CFU mL−1. Where appropriate, after growth arrest, cells were incubated with 1 μg mL−1 lysostaphin in TSB for 1 h at 37°C to partially remove the cell wall prior to the addition of daptomycin.
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