|
| Status |
Public on Jul 03, 2013 |
| Title |
LUNG_PR8A_d03_553_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
mouse lung, H1N1 influenza A PR8 (0.2LD50), 3 days
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6 (JAX 664)
age: 5-8 weeks
Sex: male
tissue: whole lung
perturbation: H1N1 influenza A PR8 (0.2LD50)
time post-infection: 3 days
mouse id: 553
|
| Treatment protocol |
5- to 8-week-old male C57BL/6 were intranasally infected with different doses of H1N1 influenza A viruses or sham infected under light isoflurane anesthesia. Infectious doses were supplied in a volume of 50ul sterile pharmaceutical-grade physiological saline.
Lungs were harvested at various time points post infection. Immediately after euthanasia, lungs were perfused in situ via the right cardiac ventricle with PBS/EDTA (10mM) pre-warmed to 37°C and then immediately processed for down-stream applications.
Entire lungs were immediately transferred into 1.5ml RNAlater (Ambion), chopped into small pieces, left at 4°C overnight and then placed at -20°C for prolonged storage.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Preserved whole lung pieces were homogenized in Trizol and total RNA was extracted, followed by clean-up and DNase I treatment with the QIAGEN RNeasy kit in accordance with the prescribed protocol provided with the kit. Quality control was performed with the Agilent Bioanalyser.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cDNA were prepared after amplification with the Ovation RNA Amplification System V2 (NuGEN Technologies).
5ug of the amplified product was used for UNG-based abasic site abrasion and biotinylation according to NuGen's Illumina Solution application notes 2.
|
| |
|
| Hybridization protocol |
2ug of the biotinylated sample was used for hybridization.
Hybridization conditions were adjusted according to NuGen's Illumina Solution application notes 2, or else the standard Illumina hybridization protocol was followed.
|
| Scan protocol |
Standard Illumina scanning protocol.
|
| Description |
4270243042_D
0.2LD50PR8d3
|
| Data processing |
The data were normalised by quantile normalisation using the GeneSpring GX 10.0 analysis platform (Agilent), log2 transformed and median baseline transformed.
|
| |
|
| Submission date |
Nov 29, 2012 |
| Last update date |
Jul 03, 2013 |
| Contact name |
Marlene Brandes |
| Organization name |
NIH, NIAID
|
| Department |
Laboratory of Systems Biology
|
| Lab |
Lymphocyte Biology Section
|
| Street address |
9000 Rockville Pike
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
| |
|
| Platform ID |
GPL6887 |
| Series (2) |
| GSE42638 |
Identification of biological processes that distinguish lethal from non-lethal influenza infection |
| GSE42641 |
A Top-down Systems Analysis Identifies an Innate Feed-forward Inflammatory Circuit Leading to Lethal Influenza Infection |
|
