|
| Status |
Public on Aug 28, 2013 |
| Title |
2.P.D |
| Sample type |
RNA |
| |
|
| Source name |
2.Patient.Diluent
|
| Organism |
Homo sapiens |
| Characteristics |
challenge: Diluent
diagnosis: P
subject: 2
cell type: CD4+ T-cells
|
| Treatment protocol |
PBMCS were cultured with grass pollen extract or diluent.
|
| Growth protocol |
PBMCs were prepared from fresh blood by means of centrifugation on Ficoll-Hypaque, washed, and stimulated with grass pollen extract (ALK-Abelló A/S; 100 μg/mL) at a density of 10^6 cells/mL and in a total volume per well of 2 mL of cell culture medium or diluent (PBS). After 7 days of incubation at 37°C and 5% CO2
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total CD4+ T cells were purified from PBMCs by positive sorting using anti-CD4-coated paramagnetic microbeads and a MACS system (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany).
|
| Label |
biotin
|
| Label protocol |
RNA extraction was performed using a miRneasy Mini Kit (Qiagen). For each sample, 250 ng of total RNA was amplified and labeled using an Illumina TotalPrep 96 RNA Amplification Kit as per the manufacturer’s instructions. The quantity of the labeled cRNA was measured using a NanoDrop spectrophotometer, and the quality and size distribution of the labeled cRNA assessed using an Agilent 2100 Bioanalyzer.
|
| |
|
| Hybridization protocol |
The hybridization, staining, washing and scanning was performed as per the manufacturer’s instructions.
|
| Scan protocol |
The hybridization, staining, washing and scanning was performed as per the manufacturer’s instructions.
|
| Description |
X2D
mRNA profiling
|
| Data processing |
Gene expression microarrays were analyzed using the R statistical programming language using functions of the gene expression microarray analysis package 'lumi'.
|
| |
|
| Submission date |
Aug 27, 2013 |
| Last update date |
Aug 28, 2013 |
| Contact name |
Colm Eamonn Nestor |
| E-mail(s) |
colm_nestor@hotmail.com
|
| Organization name |
Linkoping University
|
| Street address |
KEF, Hälsuniversitetet
|
| City |
Linkoping |
| ZIP/Postal code |
581 85 |
| Country |
Sweden |
| |
|
| Platform ID |
GPL6884 |
| Series (1) |
| GSE50223 |
Allergen challenged CD4+ cells from allergic patients and healthy controls |
|
