|
| Status |
Public on Apr 18, 2016 |
| Title |
Fly_NoListeria_AmpicillinTreatment_216hours_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Whole fly, Uninfected, Ampicillin treatment, 216 hours post infection
|
| Organism |
Drosophila melanogaster |
| Characteristics |
listeria infection status: Uninfected
ampicillin treatment: Ampicillin treatment
tissue: whole fly
gender: male
age: adult 5-7 day post-eclosion
strain: w1118 (Bloomington Stock number 6326)
|
| Treatment protocol |
At 5 to 7 days post eclosion male flies were anesthetized with carbon dioxide and were injected with 50nl of Listeria (OD600 = 0.001, or approximately 100CFUs) or left manipulated. Following injection, flies were placed in vials containing dextrose fly media and incubated at 29°C. Samples were separated into three groups: Moribund, recovering and uninfected control. Moribund flies were infected and samples were collected on the following days post infection (dpi): 0.25, 1, 2, 2.25, 3, 4, 5 and 6. Recovering flies were infected and flipped onto dextrose fly media containing 1 mg/ml ampicillin 2 days post infection. Recovering samples were collected on the following dpi: 2.25, 3, 4, 5, 6, 7, 9, and 16. Uninfected control flies were not infected but were flipped onto dextrose fly media containing 1mg/ml ampicillin 2 dpi. Uninfected control samples were collected on the following dpi: 2.25, 3, and 9.
|
| Growth protocol |
Flies were maintained on standard dextrose media at 25°C with 65% humidity and 12 hour light/dark cycles. Shortly after eclosion, adult flies were collected into bottles containing dextrose fly media. At least 24hours post eclosion adult flies were anesthetized with carbon dioxide, males were sorted into groups of 20 and placed into vials containing standard dextrose fly media. Experiments were performed on male flies 5-7 days post eclosion.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
For each sample 20 male flies were anesthetized with carbon dioxide, collected into TRIzol, homogenized and stored at -80°C. RNA preparation was preformed using a standard TRIzol RNA protocol.
|
| Label |
Cy3
|
| Label protocol |
Samples were labeled using the Quick Amp Labeling Kit, One-Color Agilent (5190-0442) following the manufacture's protocol.
|
| |
|
| Hybridization protocol |
Samples were hydrized using the Agilent Gene Expression Hybridization Kit (5188-5242) following the manufacture's protocol.
|
| Scan protocol |
Samples were scanned using a Agilent Technologies Scanner G2505C and feature intensities were determined by Agilent's Feature Extraction software(v10.7.3.1) using grid 018972_D_F_20110615.
|
| Data processing |
We used Genespring v12.1 (Agilent) for normalization and baseline transformed the data. Microarrays were normalized to the 75th percentile. The median expression level on 0 dpi was set as the baseline.
|
| |
|
| Submission date |
Sep 02, 2014 |
| Last update date |
Apr 18, 2016 |
| Contact name |
Alexander Louie |
| Organization name |
Stanford University
|
| Department |
Microbiology and Immunology
|
| Lab |
Schneider
|
| Street address |
299 Campus Dr
|
| City |
Stanford |
| State/province |
CA |
| ZIP/Postal code |
94305 |
| Country |
USA |
| |
|
| Platform ID |
GPL7300 |
| Series (1) |
| GSE60985 |
Transcriptional profiling of Drosophila during Listeria infection as they die and as they recover upon ampicillin treatment |
|
