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| Status |
Public on Mar 29, 2016 |
| Title |
Host response to systemic bacterial challenge (sepsis) with E. coli strains CFT073 and F11 at 6 and 12 hours post inoculum |
| Organism |
Danio rerio |
| Experiment type |
Expression profiling by array
|
| Summary |
Host response to systemic bacterial challenge (sepsis) with E. coli strains CFT073 and F11 at 6 and 12 hours post inoculum
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| Overall design |
48 hpf embryos were manually dechorianated, briefly anesthetized with 0.77 mM ethyl 3-aminobenzoate methanesulfonate salt (tricaine) (Sigma-Aldrich), and embedded in 0.8% low melt agarose (MO BIO Laboratories) without tricaine. After the agarose solidified, embryos were immersed in E3 media lacking methylene blue. Prior to injection, 1 ml of bacterial culture was pelleted, washed with 1 ml sterile PBS, and re-suspended in PBS to obtain ~1X109 CFU/ml. PBS. One nl of this bacterial suspension containing ~1000 CFU was microinjected into the bloodstream via the circulation valley using an Olympus SZ61 or SZX10 stereomicroscope together with a YOU-1 micromanipulator (Narishige), a Narishige IM-200 microinjector, and a JUN-AIR model 3-compressor. For each experiment, the average CFU per injection was determined by adding 10 1-nl drops to 1 ml of 0.7% NaCl, which was then serial diluted and plated on Luria-Bertani (LB) agar plates. Mock-infected controls were inoculated with 1 nl sterile PBS. Following injection, embryos were removed from agar and placed individually into wells of a 48-well plate (Nunc) containing E3 medium and incubated at 28.5°C. Experiments were performed in biological quadruplicate.
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| Contributor(s) |
Mulvey MA, Barber AE |
| Citation(s) |
27303721 |
| |
| Submission date |
Mar 28, 2016 |
| Last update date |
Jun 28, 2016 |
| Contact name |
Amelia E Barber |
| Organization name |
Leibniz-HKI
|
| Department |
Fungal Informatics
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| Street address |
Adolf-Reichwein-Straße 23
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| City |
Jena |
| State/province |
Thuringia |
| ZIP/Postal code |
07745 |
| Country |
Germany |
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| Platforms (1) |
| GPL13390 |
Agilent-021626 Danio rerio LU/IBL_D.rerio_44k (Feature Number version) |
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| Samples (16)
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| Relations |
| BioProject |
PRJNA316620 |
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