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One milliliter from each culture was added to 20 ml LB broth and incubated for 4 h at 120 rpm.
One milliliter from each of the cultures was inoculated into a 250 mL flask, containing 50 mL of YPD medium, and incubated for 5 hours under the above conditions.
One milliliter from each tube was collected at 0, 6, 12, and 24 h and immediately heat killed at 60°C for 30 min and stored at −20°C before testing with the test strip.
Model time-kill curves were determined by plotting mean colony counts (log10 CFU per milliliter) from each model versus time.
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Three milliliters from each supernatant fraction was collected and evaporated under a gentle nitrogen stream.
Two milliliters from each water/glycerol mixture were stacked on top of each other, starting with the 30% mixture at the bottom of the tube.
One-milliliter liquor from each reactor was used to inoculate liquid Luria Bertani (LB) medium containing the investigated heavy metal (sterilized by filtration) in that reactor.
One milliliter from the top was collected and analyzed by immunoblotting.
A five-parameter regression formula was used to calculate the sample concentrations (picograms per milliliter) from the standard curves.
Hydrolysis conditions were 60 min of hydrolysis time, 50°C of temperature, pH 8.0, and a specific activity of 0.3 AU per milliliter from Alcalase.
Two milliliter were taken from each sample, equal amount of ACN was added and then strongly shaken for 1 min and the mixture was stored at 4 °C overnight.
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