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. 2019 Jan 10;14(1):e0210534. doi: 10.1371/journal.pone.0210534

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© 2019 Heller, Spence

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 1 — The overall assay procedure involves the growth of a single colony of bacteria overnight in selective media at 37°C with shaking (not shown). The OD600 is measured and a diluted culture of 10 mL is prepared by dilution to an OD600 of approximately 0.005. After 2 hours of growth at 37°C, 1 mL of the diluted culture is removed and the remainder of the diluted culture is mixed with the antibiotic to which the bacteria are susceptible. Aliquots are removed at 20, 40, and 60 minutes after adding the antibiotic for OD600 determination; next, each aliquot is centrifuged at 30,000g for 30 seconds. The supernatant is transferred to a black bottom 96 well plate for the quantitative determination of ATP.