|
| Status |
Public on Dec 01, 2010 |
| Title |
Ferret Blood, uninfected, on Day 0, biological rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Ferret
|
| Organism |
Mustela putorius furo |
| Characteristics |
tissue: Blood
virus strain: Uninfected
|
| Treatment protocol |
In experiments with SARS-CoV infection, intranasal infection of ferrets with 10^3 TCID SARS-CoV / Tor2 strain was performed. Ferrets were euthanized on day 0 (before infection), day 1 and day 2 (post infection). Peripheral blood was collected in PAXgene tubes ((BD®) and lung tissue was excised and put in RNAlater (Ambion, Austin, TX) immediately. In experiments with IFN-a2b, subcutaneous injection of ferrets with 1 microgram/kilogram of IFN-a2b was performed on the back of ferrets. Ferrets were euthanized on day 0 (before injection), day 1 and day 2 (post injection). Peripheral blood was collected in PAXgene tubes and lung tissue was excised and put in RNAlater (Ambion, Austin, TX) immediately.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from blood was extracted with PAXgene Blood RNA Kit (Qiagen) in accordance with the prescribed protocol provided with the kit. Total RNA from lung was extracted with TRIzol (Invitrogen) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix Two-Cycle protocol from 100 ng total RNA (Expression Analysis Technical Manual, 2004, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Canine Genome 2.0 Array in Affymetrix Hybridization Oven 640. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000 7G
|
| Description |
Gene expression data from ferret blood, uninfected
|
| Data processing |
The data were analyzed with PLIER using Expression Console Version 1.1 (Affymetrix) default analysis settings.
|
| |
|
| Submission date |
Jun 25, 2010 |
| Last update date |
Dec 01, 2010 |
| Contact name |
Longsi Ran |
| E-mail(s) |
lran@uhnresearch.ca
|
| Phone |
416-581-7607
|
| Organization name |
Toronto General Research Institute
|
| Department |
Division of Experimental Therapeutics
|
| Lab |
Dr. David J Kelvin
|
| Street address |
101 College Street
|
| City |
Toronto |
| State/province |
ON |
| ZIP/Postal code |
M5G 1L7 |
| Country |
Canada |
| |
|
| Platform ID |
GPL3738 |
| Series (1) |
| GSE22581 |
Early gene expression events in ferrets in response to SARS coronavirus infection versus direct interferon-alpha2b stimulation |
|
