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OSM shall limit any action taken without such notice and opportunity to a temporary suspension for a maximum term of 90 days pending a decision on a final suspension, revocation or other action after such notice and opportunity have been provided.
A final suspension was made in complete RPMI 1640 (Hyclone), supplemented with 10% [v/v] FBS, 100 U/ml penicillin, 100 µg/ml streptomycin, 0.25 µg/ml and fungizone.
The day of the test, L. pneumophila cells from fresh BCYE agar plates (4 days of growth at 30°C) were resuspended in sterile water and adjusted to a final suspension with an OD600nm of 5 . 10µl of the bacterial suspension was added into each well of the plate with different dilutions of the drugs.
After temperature equilibration, the ionophore A23187 was added to a final suspension concentration of 10 μM.
Four rinse cycles were performed with Millipore water, resulting in a final suspension of the particles in 50 μL of water.
A final suspension of single cells was prepared in ice-cold PBS containing 0.5% BSA and 2mM EDTA (MACS Buffer) for subsequent spermatogonial stem cell enrichment.
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Note the lower OD (∼0.6) in plates was due to a decreased path length of the 60 µl final suspension, in a well, compared to a 1 cm path length in a spectrophotometer.
DNA concentration in the nuclei suspension was determined by A260 measurement (Nanodrop) and a final DNA suspension between 1.2 and 1.6 mg/mL was prepared by dilution with CaCl2 added to a final concentration of 1 mM.
Sorption experiments were initiated by adding 20 mL of the final suspension to a 100-mL polypropylene bottle containing 0.8 mL to 16 mL of 15-mM Cu(NO3 2 solution.
The AgNP suspension was then centrifuged (2200 g, 15 min) producing a final stock suspension with an average Ag concentration of 35.7 mg Ag L-1 (SD = 5.6 mg Ag L-1, n = 11), 8% of which was dissolved ionic Ag+[43].
The total sperm count was assessed in the final suspension by using a hemacytometer.
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