|
| Status |
Public on Dec 31, 2006 |
| Title |
proband 1 Tcells, before exercise |
| Sample type |
RNA |
| |
|
| Source name |
human peripheral T-lymphocytes
|
| Organism |
Homo sapiens |
| Characteristics |
human peripheral T-lymphocytes
|
| Treatment protocol |
8ml of whole blood were collected in BD Vacutainer CPT tubes. 400 µl of RosetteSep Tcell Enrichment cocktail were added immediately following the blood withdrawal. Tubes were inverted five times and incubated for 20 minutes at room temperature. Tubes were next centrifuged for 20 minutes at 1850g at room temperature. The cells were washed two times in icecold (4°C) PBS/ 2mM EDTA once in a volume of 50ml and the second time with a final volume of 5ml, spun down for 10 minutes at 500g at 4°C, the supernatant was removed by vacuum suction and the final pellet was lysed with Trizol.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted from Trizol through addition of chloroform and phase separation using PLG tubes. Additionally RNA was cleaned up using the Rneasy Micro Kit with a Dnase step.
|
| Label |
biotin
|
| Label protocol |
biotinylated RNA was prepared from 3 µg total RNA according to the Affymetrix manual (2003)
|
| |
|
| Hybridization protocol |
6.5 µg biotinylated RNA were hybridised for 16h at 45°C on geneChip human U133 A 2.0 arrays. GeneChips were washed and stained in the Affymetrix Fluidics station 400.
|
| Scan protocol |
GeneChips were scanned with the Affymetrix Scanner 3000.
|
| Description |
proband 1 Tcells, before exercise
|
| Data processing |
The data were first analysed with Affymetrix GCOS 1.2 using default analysis settings.
|
| |
|
| Submission date |
Aug 14, 2006 |
| Last update date |
Oct 17, 2006 |
| Contact name |
Sandy Mosig |
| E-mail(s) |
sandy.mosig@mti.uni-jena.de, sandy.mosig@googlemail.com
|
| Phone |
+49-3641-934813
|
| Fax |
+49-3641-933950
|
| Organization name |
University Hospital Jena
|
| Department |
Molecular Hemostaseology - Institute of Vascular Medicine
|
| Street address |
Bachstrasse 18
|
| City |
Jena |
| ZIP/Postal code |
07743 |
| Country |
Germany |
| |
|
| Platform ID |
GPL571 |
| Series (1) |
| GSE6053 |
Gene Expression analyses in PBMCs and T-lymphocytes following acute exhausting exercise |
|
