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Growth experiments

MW Michaela Wenzel
MD Marien P. Dekker
BW Biwen Wang
MB Maroeska J. Burggraaf
WB Wilbert Bitter
JW Jan R. T. van Weering
LH Leendert W. Hamoen
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B. subtilis 168 was aerobically grown in LB. Overnight cultures were diluted to an OD600 of 0.05 and allowed to grow until an OD600 of 0.4 prior to addition of antibiotics at different MIC multiples. Growth was monitored for 8 h using a Biotek Synergy MX plate reader equipped with Gen5 software. Concentrations leading to a reduced growth rate without causing massive cell lysis within the first 30 min of antibiotic exposure were chosen for electron microscopy. Daptomycin requires the presence of 1.25 mM CaCl2. Addition of CaCl2 did not affect growth of B. subtilis. Experiments were performed in duplicates.

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