5. To conduct an endpoint CFU/mL count, serially dilute
unsealed culture in sterile PBS. Spread out a minimum aliquot
of 50 μL of each dilution. Incubate in a static incubator at
37
C with 5% CO 2 (see Note 15). After approximately
Fig. 2 Plate maps of high-throughput screening varieties. (a) Single-shot highthroughput screen. (b) Dilution series of multiple compounds, ranging from high
to low concentration. (c) Checkerboard assay, where dilution series of two
different drugs are assayed against each other, which simultaneously shows
individual activities as well as synergistic or antagonistic activities of combined
antimicrobial compounds [16]. In all plate maps, there is a perimeter ring of
dH 2 O (light blue) to prevent evaporation around the plate and row 11 contains
culture only (yellow) and media only (gray) control wells
In vitro Drug Screening of Hypoxic NRP Mycobacteria
255
unsealed culture in sterile PBS. Spread out a minimum aliquot
of 50 μL of each dilution. Incubate in a static incubator at
37
C with 5% CO 2 (see Note 15). After approximately
Fig. 2 Plate maps of high-throughput screening varieties. (a) Single-shot highthroughput screen. (b) Dilution series of multiple compounds, ranging from high
to low concentration. (c) Checkerboard assay, where dilution series of two
different drugs are assayed against each other, which simultaneously shows
individual activities as well as synergistic or antagonistic activities of combined
antimicrobial compounds [16]. In all plate maps, there is a perimeter ring of
dH 2 O (light blue) to prevent evaporation around the plate and row 11 contains
culture only (yellow) and media only (gray) control wells
In vitro Drug Screening of Hypoxic NRP Mycobacteria
255