|
| Status |
Public on Dec 12, 2007 |
| Title |
WT S49_untreated_2h, biological rep1 |
| Sample type |
RNA |
| |
|
| Source name |
S49 WT cells 2h treatment with 8-CPT-cAMP
|
| Organism |
Mus musculus |
| Characteristics |
Balb/C, Lymphoma, tumor
|
| Treatment protocol |
cells were treated with 100 mM 8-CPT-cAMP for the indicated times
|
| Growth protocol |
D- S49 cells were grown in suspension cultures with Dulbecco's modified Eagles' medium supplemented with 10% heat-inactivated horse serum and 10 mM HEPES in a humidified atmosphere containing 10% CO2 at 37ºC. Cultures were initiated at a density of 2.5-5 x 10E5 cells/ml and carried out for the indicated times.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total cellular RNA was isolated from cells by using RNeasy mini-columns
|
| Label |
biotin
|
| Label protocol |
Total cellular RNA was reverse transcribed with superscript II reverse transcriptase and an oligo-dT primer containing a T7 RNA polymerase promoter. Single stranded cDNA was converted into double-stranded cDNA (ds-cDNA) and then purified (MessageAMP aRNA Kit, Ambion, Austin, TX). Biotinylated cRNA was generated from ds-cDNA by in vitro transcription (IVT) (MessageAMP aRNA Kit, Ambion). After a further round of purification, IVT reactions yielded 30–70 µg of biotinylated cRNA, which was fragmented to ~100–base pair fragments before hybridization.
|
| |
|
| Hybridization protocol |
Each Affymetrix 430 A chip was hybridized to 10 µg of fragmented cRNA.
|
| Scan protocol |
Arrays were hybridized and scanned with a GeneArray Scanner (Hewlett-Packard/Affymetrix)
|
| Description |
2h 8-CPT-cAMP treated WT S49 cells, biological rep1
|
| Data processing |
Cel files were converted to signal values by gcRMA
|
| |
|
| Submission date |
Dec 11, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
Alexander C Zambon |
| Organization name |
Keck Graduate Institute
|
| Department |
Biopharmaceutical Sciences
|
| Lab |
Zambon
|
| Street address |
517 Watson Drive
|
| City |
Claremont |
| State/province |
CA |
| ZIP/Postal code |
91711 |
| Country |
USA |
| |
|
| Platform ID |
GPL339 |
| Series (1) |
| GSE9727 |
Gene Expression in S49 Deathless (D-) cell variant |
|
