|
| Status |
Public on May 19, 2014 |
| Title |
EOS from saline lung_rep4 |
| Sample type |
RNA |
| |
|
| Source name |
eosinophils sorted from saline challenged lung
|
| Organism |
Mus musculus |
| Characteristics |
strain: BALB/c
tissue: lung
cell type: eosinophil
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Following the QIAGEN RNeasy kit procedures
|
| Label |
biotin
|
| Label protocol |
The WT- Ovation Pico RNA Amplification System Version 1.0 (Nugen) is used to prepare target cDNA. Then the WT-Ovation Exon Module v1.0 (NuGEN) is used to create target sense transcript cDNA. Finally the FL-Ovation cDNA Biotin Module V2 (NuGEN) chemically fragments and labels the target sense cDNA
|
| |
|
| Hybridization protocol |
Hybridization chamber: a standard Probe Array Cartidge (GeneChip Mouse Gene 1.0 ST Array Affymetrix). Hybridization cocktail for a single probe array that contains 2.5 ug of fragmented cRNA, 50 pM Control Oligonucleotide B2 (Affymetrix), 20X Eukaryotic Hybridization Controls (1.5 pM bioB, 5 pM bioC, 25 pM bioD, 100 pM cre) (Affymetrix), 10% DMSO, and 1X Hybridization Buffer. Heat hybridization cocktail to 99C for 2 minutes, to 45C for 5 minutes, and spin at maximum speed in a microcentrifuge for 5 minutes. Fill probe array with 90 uL of 1X Hybridization Buffer. Incubate at 45C for 10 minutes in the GeneChip Hybridization Oven 640 (Affymetrix) rotating at 60 rpm. Remove 1X Hybridization Buffer and fill probe array with 90 uL of the hybridization cocktail. Incubate at 45C for 18 hrs in the Hybridization Oven rotating at 60 rpm.
|
| Scan protocol |
Affymetrix GeneChip Scanner 3000 7G, Scanning software: Genechip Operating Software (Affymetrix)
|
| Description |
Gene expression data from sorted eosinophils
SAMPLE 4
|
| Data processing |
Raw data were analyzed by Genespring GX 11 using RMA
|
| |
|
| Submission date |
May 16, 2014 |
| Last update date |
May 19, 2014 |
| Contact name |
Marc E Rothenberg |
| E-mail(s) |
rothenberg@cchmc.org
|
| Phone |
513-636-7177
|
| Organization name |
Cincinnati Children's Hospital Medical Center
|
| Department |
Allergy
|
| Lab |
Rothenberg
|
| Street address |
3333 Burnet Ave. MLC-7028
|
| City |
Cincinnati |
| State/province |
OH |
| ZIP/Postal code |
45229 |
| Country |
USA |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE57757 |
Expression profiles of sorted murine lung Eosinophils following allergen challenge |
|
