The WT USA300 strain and codY mutant containing either pGYbrnQ1∷lux or pGYbcaP∷lux were grown to mid-late exponential phase in complete CDM and sub-cultured to a starting OD600 of 0.01 (200 uL well−1) of complete CDM in a clear bottom 96 well Microfluor 2 White Plate (Thermo). Plates were incubated at 37°C with constant shaking. Luminescence and OD600 were ready at hourly intervals on the BioTek Synergy H4 Hybrid Reader (BioTek, Winooski, VT) with 1 s of integration and a gain of 200.
Microfluor 2 white plate
Manufactured by Thermo Fisher Scientific
The Microfluor 2 White Plate is a multi-well microplate designed for fluorescence-based assays. The plate is made of a white polystyrene material that maximizes the reflection of light, enhancing the signal-to-noise ratio for improved sensitivity in fluorescence measurements.
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Lab products found in correlation
2 protocols using microfluor 2 white plate
1
Luminescence Monitoring of S. aureus
2
Characterizing MRSA Growth Dynamics
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The methicillin-resistant Staphylococcus aureus COL was used in superinfection experiments. A mutant strain of this bacterium with a deletion of seb, the gene encoding staphylococcal enterotoxin B (SEB), was previously described [37 (link)]. These strains will be simply referred to as COL and COL Δseb, respectively. For bacterial growth curve analysis, cells were picked from a tryptic soy agar (TSA) plate, grown overnight in Brain Heart Infusion (BHI) broth, and subcultured in half-BHI medium (BHI broth diluted 1:2 in sterile Milli-Q water). OD600 was adjusted to 0.05 before 200 μL/well of cultures were placed in quadruplicate in a 96-well flat-bottom Microfluor 2 White plate (Thermo Fisher Scientific). Bacterial growth was monitored at 37°C with shaking using a Biotek Synergy H4 plate reader, and OD600 readings were recorded at one-hour intervals.
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