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The donor plate was a 96-well filtrate plate (Multiscreen® IP Sterile Plate PDVF membrane, pore size is 0.45 µM, catalog no. MAIPS4510) and the acceptor plate was an indented 96-well plate (Multiscreen®, catalog no. MAMCS9610 from Millipore).
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Serial dilutions of the filtrate were plated on an indicator bacterial strain and plaque counts were determined after 16 h of incubation at 37 °C.
Filtrate was plated to ensure the absence of S. aureus (data not shown).
The filtrate was plated to determine the amount of unattached phages.
Organs were weighed separately, crushed, macerated, and suspended in sterile saline, and the filtrate was plated on MacConkey agar to determine the number of translocated bacteria.
The salt making process is done by (1) collecting and socking salty soils in jerry cans, (2) vigorously stirring and filtering the mixture to get salt concentrated water, (3) boiling the filtrates to make salts (Plate 1).
Samples were filtrated through 0.45 μm plate filter and loaded to HPLC for sugar determination.
The culture filtrate was confirmed by dilution plate technique to have a density of 3.6 ± 0.49 CFU/mL and a BPSA emulsion degradation activity of 0.28 U/mL.
This sample was filtered using a 0.45 μm pore sized PTFE membrane filter plate, and the filtrates were collected into a new 96-well plate.
The plates with the filtrate samples were incubated overnight at 37°C and were observed for plaque formation.
Phages in the filtrate are grown on agar plates seeded with the bacterium the engineers want to kill.
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