|
| Status |
Public on Jun 09, 2010 |
| Title |
TOL10 |
| Sample type |
RNA |
| |
|
| Source name |
Tolerant10
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Blood
phenotype: Tolerant
|
| Treatment protocol |
Participants in this study received no protocol related treatment.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted from whole blood collected in ABI Tempus tubes. Extraction was performed using ABI6100 instrument.
|
| Label |
biotin
|
| Label protocol |
Biotin-labelled target for the microarray experiment was prepared using 100 ng of total RNA and cDNA was synthesized using the GeneChip® WT (Whole Transcript) Sense Target Labelling and Control Reagents kit as described by the manufacturer (Affymetrix). The sense cDNA was then fragmented by UDG (uracil DNA glycosylase) and APE 1 (apurinic/apyrimidic endonuclease 1) and biotin-labelled with TdT (terminal deoxynucleotidyl transferase) using the GeneChip® WT Terminal labelling kit (Affymetrix, Santa Clara, USA). Hybridization was performed using 5 micrograms of biotinylated target, which was incubated with the GeneChip® Human Gene 1.0 ST array (Affymetrix) at 45°C for 16–20 hours.
|
| |
|
| Hybridization protocol |
Hybridization was performed using 5 micrograms of biotinylated target, which was incubated with the GeneChip® Human Gene 1.0 ST array (Affymetrix) at 45°C for 16–20 hours. Following hybridization, non-specifically bound material was removed by washing and detection of specifically bound target was performed using the GeneChip® Hybridization, Wash and Stain kit, and the GeneChip® Fluidics Station 450 (Affymetrix).
|
| Scan protocol |
The arrays were scanned using the GeneChip® Scanner 3000 7G (Affymetrix) and raw data was extracted from the scanned images and analyzed with the Affymetrix GeneChip Operation System (GCOS, Affymetrix).
|
| Description |
Gene expression data from Tolerant 10
|
| Data processing |
Microarray data background adjustment, normalization, and summarization were performed using the robust multichip average method.
|
| |
|
| Submission date |
Jun 08, 2010 |
| Last update date |
Feb 16, 2021 |
| Contact name |
Sai Kanaparthi |
| E-mail(s) |
[email protected]
|
| Organization name |
Immune Tolerance Network
|
| Department |
BiG
|
| Street address |
7500 Old Georgetown Rd
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20814 |
| Country |
USA |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE22229 |
Identification of a B cell signature associated with renal transplant Tolerance in humans |
|
| Relations |
| Reanalyzed by |
GSE49198 |
| Reanalyzed by |
GSE166865 |
