|
| Status |
Public on Jan 01, 2008 |
| Title |
JT_Sw3_15 |
| Sample type |
RNA |
| |
|
| Source name |
sham operated female
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6J
age: 12-14 weeks
sex: female
|
| Treatment protocol |
Mice were sham operated. Animals received buprenorphine (0.05mg/kg body weight) bid for the first 2 days following thoracotomy. All procedures were in accordance with animal welfare guidelines and were approved by the respective governmental agencies.
|
| Growth protocol |
12-14 week old male and female C57BL/6J mice were housed under climate-controlled conditions with a 12h light/dark cycle and were provided with standard food and water ad libitum.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated 14 days after surgery of the left ventricle using TRIzol Reagent (Invitrogen).
|
| Label |
biotin labeled
|
| Label protocol |
The amplification and labeling of the RNA samples were carried out according to the manufacturer's instructions (Affymetrix, Santa Clara, CA). Briefly, total RNA was quantified by UV-spectroscopy and its quality evaluated by analysis on a LabChip (BioAnalyzer, AGILENT Technologies, Santa Clara, CA). In the presence of an oligo(dT)24 primer containing a T7 RNA polymerase promoter (TIBMOL Biol, Berlin, Germany) one to three micrograms of total RNA were used for double-stranded cDNA synthesis (SuperScript transcriptase, Life Technologies, Inc., Carlsbad, CA). Labeled complementary RNA (cRNA) was prepared from double-stranded cDNA by in vitro transcription using the GeneChip RNA transcript labeling kit (Affymetrix, Santa Clara, CA). After cleanup (Qiagen, Hilden, Germany), the biotin-labeled cRNA was fragmented by alkaline treatment (40 mM Tris-acetate (pH 8.2), 100 mM potassium acetate, and 50 mM magnesium acetate) at 94°C for 35 minutes.
|
| |
|
| Hybridization protocol |
Fifteen micrograms of each cRNA sample was hybridized for 16 hours at 45°C to rat expression Affymetrix RAE 430A GeneChip arrays. Following the hybridization arrays were washed and stained with streptavidin-phycoerythrin using a fluidics station in accordance with the Affymetrix protocol.
|
| Scan protocol |
Affymetrix GeneChip System confocal scanner 2500
|
| Description |
sham operated female control mouse
|
| Data processing |
Affymetrix GeneChip Operating Software (GCOS)
|
| |
|
| Submission date |
Feb 05, 2007 |
| Last update date |
Mar 06, 2007 |
| Contact name |
Henning Witt |
| E-mail(s) |
[email protected]
|
| Phone |
++49-30-450578727
|
| Organization name |
Max Planck Institute for Molecular Genetics
|
| Department |
Hans Lehrach
|
| Lab |
Patricia Ruiz
|
| Street address |
Ihnestrasse 65
|
| City |
Berlin |
| ZIP/Postal code |
14195 |
| Country |
Germany |
| |
|
| Platform ID |
GPL339 |
| Series (1) |
| GSE6970 |
Sex-specific early response to pressure overload in mice |
|
