Infinite f nano

Overnight cultures of S. aureus were diluted to OD = 0.025 in BHI broth supplemented with 5 mM calcium chloride and antibiotics and grown for 1 h and 15 min shaking at 37°C. Cultures were normalized to the OD = 0.1 and phage ɸNM4γ4 was added to the appropriate MOI. After inverting to mix, 150 μL of each culture were added to a flat-bottom 96-well plate (Grenier 655180), and the plate was incubated at 37°C with shaking in a TECAN Infinite F Nano+ with OD600 measurements recorded every 10 min for 24 h
For Streptococcus pyogenes assays, overnight cultures of S. pyogenes were diluted back 1:10 in fresh Thy media supplemented with 5 mM calcium chloride and 2 mg/mL sodium bicarbonate and grown for 1 h and 30 min without shaking at 37°C. Cultures were normalized to the OD = 0.05 and phage A1 (and derivatives) was added to the appropriate normalized MOI. After inverting to mix, 200 μL of each culture was added to a flat-bottom 96-well plate (Grenier 655180), and the plate was incubated at 37°C without shaking in a TECAN Infinite F Nano+ with OD600 measurements recorded every 10 min for 24 h
To quantify colony forming units (CFUs) remaining at the conclusion of a liquid growth interference assay, 200 μL of each culture was serially diluted 1:10 eight times, and plated on plain BHI plates.

Measurements for absorbance (at 600 nm) and fluorescence (excitation wavelength = 485 nm; emission wavelength = 535 nm) were taken using a TECAN Infinite F Nano+. For each experimental strain, promoter activity was measured as (Fe)/(Ae) - (Fc)/(Ac) where F = fluorescence, A = absorbance, e = experimental strain and c = non-fluorescent control strain.