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For each substitution line, 300 pairs of sexually mature flies were placed in egg collecting chambers for 8 hours.
Females were placed in egg laying chambers containing standard fly food and fecundity measured by counting the total number of eggs laid over a five day period.
3-dpf embryos were placed in egg water containing 0.5 μg/ml 4-hydroxyanisole (Sigma) for 3 days and imaged.
6-dpf embryos were placed in egg water containing 1 nM melanin concentrating hormone (Sigma) for 10 minutes (Logan et al., 2006).
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By contrast, it proved hard to grow the weakened strain of swine flu that could be placed in eggs.
The surrogate egg shells containing injected and control un-injected eggs were placed in an Egg Incubator (P-008Q Bio-type; Showa Furanki Corporation, Japan) set at 39°C.
For embryo imaging, 50 male and 50 virgin female flies, <5 days old, were placed in an egg collection chamber with a grape agar plate with yeast paste.
After this seven day period all females were removed and placed in individual egg-laying chambers.
Egg masses placed in distilled water without crude antibiotic extract served as control.
Each collected egg was placed in a Petri dish.
Afterwards, the hole was sealed with hypoallergenic tape and the eggs were placed in an incubator.
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