|
| Status |
Public on Jan 26, 2009 |
| Title |
Pancreatic Islets_18h culture_2 mmol/l glucose_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Pancreatic islets, 18h culture, 2 mmol/l glucose
|
| Organism |
Rattus norvegicus |
| Characteristics |
Strain: Wistar (male)
Weight: 180-200g
|
| Treatment protocol |
After preculture, the islets were cultured 18h in serum-free RPMI medium containing 5 g/l BSA and 2, 5, 10 or 30 mmol/l glucose (1000 islets per condition).
|
| Growth protocol |
After isolation, the islets were precultured for 1 week in serum-free RPMI medium containing 5 g/l BSA and 10 mmol/l glucose. The medium was renewed after 24h, then every 48h.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Islet total RNA was extracted using the Absolutely microprep kit (Stratagene, city, CA, USA) following manufacturer’s instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol.
|
| |
|
| Hybridization protocol |
Fragmented cRNA was hybridized 16h at 45°C to a Rat Genome 230 2.0 array.
|
| Scan protocol |
After hybridization, the arrays were washed and stained with streptavidin-phycoerythrin and scanned using the Affymetrix 3000 GeneScanner.
|
| Description |
Before the experiment, the islets were precultured for 1 week in serum-free RPMI medium containing 5 g/l BSA and 10 mmol/l glucose
|
| Data processing |
Raw data were analyzed using the Affymetrix GCOS software. Signal intensities were scaled using the global scaling method of the statistical expression algorithm taking 150 as target intensity value. Scaling factors were less than 3 times different from each other in all used arrays.
|
| |
|
| Submission date |
Sep 17, 2008 |
| Last update date |
Jan 26, 2009 |
| Contact name |
Jean-Christophe Jonas |
| E-mail(s) |
[email protected]
|
| Phone |
+3227649575
|
| Fax |
+3227645532
|
| Organization name |
UCL, Faculty of Medicine
|
| Department |
Physiology and Pharmacology
|
| Lab |
Endocrinology and Metabolism
|
| Street address |
Av. Hippocrate, 55
|
| City |
Brussels |
| ZIP/Postal code |
B-1200 |
| Country |
Belgium |
| |
|
| Platform ID |
GPL1355 |
| Series (1) |
| GSE12817 |
Cluster analysis of rat pancreatic islet gene mRNA levels after culture in low, intermediate and high [glucose] |
|
