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Merck

Protocol for a Binary Choice Feeding Assay Using Adult, Axenic Drosophila.

STAR protocols (2020-12-31)
Bernard Charroux, Julien Royet
RESUMEN

This protocol is designed to prepare adult axenic Drosophila before monitoring their behavior in a two-choice feeding assay, where flies are confronted with an axenic versus a dead or alive bacteria-contaminated feeding solution. Several aspects of the procedure, including raising and aging flies in axenic conditions, starving adult flies, and composing feeding solutions, are detailed. The bacterium used in this protocol, Erwinia carotovora carotovora-15 2141 (Ecc-15 2141 ), is commonly used to decipher the mechanisms controlling host-pathogen interactions in the Drosophila model. For complete details on the use and execution of this protocol, please refer to Charroux et al. (2020).

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Sigma-Aldrich
Caldo LB (Lennox), Highly-referenced microbial growth powder medium, low salt, suitable for salt-sensitive E.coli culture.
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Rifampicina, ≥95% (HPLC), powder or crystals
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Spectinomycin dihydrochloride pentahydrate, powder, BioReagent, suitable for cell culture
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Caldo LB con agar (Lennox), Highly-referenced microbial growth powder medium with Agar, low salt, suitable for salt-sensitive E. coli culture.
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Sacarosa, ≥99.5% (GC), BioReagent, suitable for cell culture, suitable for insect cell culture
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Erythromycin, BioReagent, suitable for cell culture
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Tetracycline, 98.0-102.0% (HPLC)