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An advantage of this assay is that it can be initiated using a predefined number of mating pairs in a single genetic cross, which generates both control and experimental animals (expressing APP695) that will be raised under identical conditions (in the same vials of medium) and that can be readily distinguished by genetic markers.
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Larvae of the desired genotype were selected at 56 64 h AEL and transferred into vials of cornmeal medium lacking agar.
Newly-eclosed adults were collected and aged in vials of standard medium at 25°C (≤11 flies of the same sex per vial) for 5-6 days before testing.
The ability of Group II C. botulinum strains to grow at refrigeration temperatures or in media containing different NaCl concentrations was tested using vials of PYGS medium inoculated with spores of appropriate strains.
On receipt at the laboratory, fresh human blood samples were examined by darkfield microscopy for viable Borrelia and, if detected, 200 μL of blood was injected into 10-mL vials of BSK-H medium (14 ) and into 10-week-old ICR mice by the intraperitoneal route.
For a concentration of 3 vol% oxygen 22 ml of sterile air was injected into closed anoxic serum vials containing 41 ml of medium.
The isolated PBMCs were seeded in plastic vials at a density of 1 × 10 viable cells per vial in 1 ml of medium (RPMI 1640 containing 10%% fetal bovine serum, 2 mM L-glutamine and 1 % of antibiotic-antimycotic solution).
Following the addition of medium, the vials were sealed using silicone stoppers and aluminum crimp seals (Supelco), and autoclaved.
Here, flies were kept individually in 10 × 3 cm ∅ vials, provided with 5 ml of medium.
Due to the different volumes of medium within the vials the weight ratios of substrate to oxygen were almost identical at 3 and 6 vol% O2, but approx. three times less compared to vials with 12 vol% O2 and six times less compared to vials with 21 vol% initial oxygen concentration.
Media were prepared in the shape of a slope (using 5 ml of medium) inside 20 ml sterile vials designed for headspace (HS) sampling.
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