|
| Status |
Public on May 03, 2011 |
| Title |
adrenal_medulla_sham_3 |
| Sample type |
RNA |
| |
|
| Source name |
adrenal medulla after sham
|
| Organism |
Mus musculus |
| Characteristics |
gender: male
strain: C57/Bl6
|
| Treatment protocol |
Adrenal glands were frozen in liquid nitrogen. 12 µm cryosections were dehydrated in xylene. Adrenal medullary tissue was isolated by microdissection with a Leica AM6000 inverted microscope (Leica Microsystems, Wetzlar, Germany). A MicroChisel (Eppendorf, Hamburg, Germany) was used for microdissection. Tissue was aspirated in xylene. Next, xylene was evaporated (Concentrator 5303, Eppendorff, Hamburg, Germany)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from both adrenal medulla of one animal was extracted using the RNeasy Micro Kit (Qiagen, Hilden, Germany).
|
| Label |
biotin
|
| Label protocol |
Double stranded cDNA was synthesized (GeneChip® Expression 3' Amplification Two-Cycle Target Labeling and Control Reagents, 900494) and biotin labeled cRNA was in vitro transcribed (GeneChip® Expression 3' Amplification Two-Cycle Target Labeling and Control Reagents, 900494). Cleanup of the cDNA and the fragmented cRNA was carried out with the GeneChip Sample Module (Affymetrix,900371). The integrity of the total RNA and of the subsequent preparation steps was validated using the Agilent 2100 bioanalyzer (Waldbronn, Germany).
|
| |
|
| Hybridization protocol |
10 µg of fragmented cRNA were hybridized (16h) to the array. Staining and washing was conducted according to the Affymetrix protocol.
|
| Scan protocol |
The processed array was scanned using a GeneChip Scanner 3000 7G (Affymetrix).
|
| Data processing |
Data was analyzed using ArrayAssist 5.0 software (Stratagene). Probe levels were calculated using the GCRMA algorithm.
|
| |
|
| Submission date |
May 14, 2010 |
| Last update date |
May 03, 2011 |
| Contact name |
Ralf Gilsbach |
| E-mail(s) |
[email protected]
|
| Organization name |
Institute of Pharmacology
|
| Lab |
Prof. Dr. Lutz Hein
|
| Street address |
Albertstr. 25
|
| City |
Freiburg |
| ZIP/Postal code |
79104 |
| Country |
Germany |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE21829 |
Differential gene expression in adrenal medulla after cardiac pressure overload |
|
