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Aliquots of 0.1 ml were sampled at regular intervals and supplemented with 1 μl 0.1% Calcofluor (which reacts to cellulose in the cyst wall).
Aliquots of 1.5 mL were sampled over 160 min at 20-min intervals.
Aliquots of 0.2 mL were sampled from the beaker at predetermine intervals.
Suspension aliquots of 10 ml were sampled at 2, 4, 22 and 24 h incubation, respectively, and immediately analyzed on the continuous flow analyzer to determine their nitrate concentrations.
The perfusions were run for four hours, and 6 mL were sampled at t = 0 from the maternal circulation and from both reservoirs before addition of the compounds and at 2, 30, 60, 130, 190 and 240 minutes after addition of the compounds.
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Culture (5 mL) was sampled after 2 days, vortexed, and spun at 3000 × g for 5 min.
The culture medium (0.5 mL) was sampled per day and centrifuged at 4°C, 10,000 rpm for 5 min. Cell pellet and supernatant were separated.
Total bone marrow (around 5 ml) was sampled after the last boost to isolate RNA using Tri Reagent (Molecular Research Center Inc., Cincinnati, USA) according to the manufacturer's instructions.
Blood (3 mL) was sampled from every patient.
For milk, 10.0 ml was sampled and its pH was lowered to 4.6 or below.
At each water change, 500 mL was sampled and placed in a precleaned glass amber jar for testing.
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