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Before assay, the salivary sample was centrifuged for 5 minutes at 2500 revolutions per minute.
Before assay, all samples were thawed to room temperature and mixed by gentle swirling or inversion.
Before assay, 5 μl culture from each sample were diluted 200x in 0.22 μm filtered DI water (pH 7).
Before assay replacement, samples were tested with both techniques and results were comparable (A. Onderdonk, pers. comm., 2008).
Before assay, 1 ml portions of controls and samples were extracted twice in 4.5 ml of diethyl ether.
Before assay the stock was diluted into PBS by a factor of 60, which was used as working solution for luminescence.
Before assay, the organic filtrates were concentrated in vacuo in a rotary evaporator at 50°C while the aqueous extracts were lyophilized.
The separated plasma was stored frozen at −20 °C before assay.
All the solutions for activity assay were balanced in an anaerobic chamber filled with 100% N2 for 30 min before assay.
Cells were seeded at a cell density of 104 cells/well in a 96-well plate and grown for 48 h before assay.
Virions were normalized for equivalent RT activity before assay.
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