3. Drug aliquots. Store at À20
C in single use aliquots
(10–20 μL).
4. 96- or 384-well clear, flat-bottom multiwell plates.
5. Universal plate lid.
2.3 Data Analysis
1. MATLAB, R, Python, Microsoft Excel, or other scientific computing software.
3 Methods
3.1 Culturing
Bacteria
1. Inoculate M. tuberculosis in 10 mL of 7H9 broth medium in
30 mL inkwell with a starting OD 600 of ~0.05 (see Note 1).
2. Incubate in a shaking incubator (37
C, 100 rpm) for approximately 3 days to achieve log-phase culture.
3. Subculture once to OD 600 of 0.05 and incubate for another
3 days to achieve log-phase culture again.
4. Dilute the culture to OD 600 of 0.05 with fresh medium for
drug treatment in 384-well plates.
3.2 Dose Centering
The goal of this step is to determine the dose response of each
individual compound of interest in the context of the specific
growth conditions and timing of the assay. In practice, if the minimal inhibitory concentration (MIC) has already been determined
Fig. 2 DiaMOND experiment and analysis workflow
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