|
| Status |
Public on Sep 27, 2006 |
| Title |
Oxyntic mucosa_Octreotide LAR_21-days_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Oxyntix mucosa, Octreotide LAR, 21 days
|
| Organism |
Rattus norvegicus |
| Characteristics |
Sprague-Dawley, female, 8 weeks. Tissue: scraped off oxyntic mucosa.
|
| Treatment protocol |
Rats housed in wire-mesh cages at 20-24°C with constant humidity and a 12:12 h light-dark cycle, and fed ad libitum with a commercial rat diet and tap water. Overnight (one night) fast the animals were anesthetised with 0.4 ml per 100 g body weight of a combination of (per ml) 0.02 mg fentanyl, 0.02 mg haloperidol and 2 mg midazolam. Eight animals received the LAR vehicle in corresponding volume to 5 mg/animal of Octreotide LAR.
|
| Growth protocol |
Tissue were put into liquid nitrogen at once after separation from the rats.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Extraction of total RNA were done using RNeasy Midi Kit (Qiagen, Vanlencia, CA)
|
| Label |
Biotin
|
| Label protocol |
Biotin-labeled cRNA synthesis: IVT Labeling Kit (Affymetrix)
Biotin-labeled cRNA clean up: GeneChip® Sample Cleanup Module (Affymetrix).
|
| |
|
| Hybridization protocol |
Hybridization was performed in accordance with the standard array format protocol using 15µg fragmented biotinylated cRNA. The hybridization cocktails were applied to the arrays and left to hybridize at 45°C for 16 hours in an Affyetrix GeneChip Hybridization oven 640 with constant rotation (60rpm).
The chips were washed and stained using Affymetrix Fluidic Station 400 controlled by MicroArray Suite 5.0. The antibody amplification protocol EukGE-WS2v4 (Affymetrix) was used for controlling the washing and staining. Staining was performed using streptavidin phycoerythrin (SAPE, Molecular Probes, Eugene, Oregon, USA), followed by adding biotinylated antibody (Vector Laboratories) to streptavidin and then one more time with streptavidin phycoerythrin.
|
| Scan protocol |
The arrays were scanned using Agilent GeneArray Scanner.
|
| Description |
RNA from eigth animals (controls that received the LAR-vehicle) were pooled to produce a control affymetrix chip. Six animals (treatmentgroup) were given octreotide LAR and RNA from two and two animals were pooled to produce tree affymetrix chips to compare to the controlchip.
|
| Data processing |
Scanner controlled by MAS.
|
| |
|
| Submission date |
Sep 26, 2006 |
| Last update date |
Sep 26, 2006 |
| Contact name |
Sten Even Erlandsen |
| E-mail(s) |
[email protected]
|
| Phone |
004799526568
|
| Organization name |
NTNU
|
| Department |
Department of Cancer Research and Molecular Medicine
|
| Street address |
Olav Kyrres gate 9, 5 etg.
|
| City |
Trondheim |
| ZIP/Postal code |
7489 |
| Country |
Norway |
| |
|
| Platform ID |
GPL341 |
| Series (1) |
| GSE5910 |
Octreotide LAR and the oxyntic mucosa |
|
