|
| Status |
Public on Jul 06, 2011 |
| Title |
Mesentery_Ethinyl estradiol_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Mesenteric arteries, treated with ethinyl estradiol
|
| Organism |
Rattus norvegicus |
| Characteristics |
strain: Sprague Dawley
age: 9 weeks
gender: female
tissue: mesenteric artery
agent: ethinyl estradiol
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The mesenteric arterial tree was dissected free from veins and adipose tissue under RNAlater (Ambion). The RNAlater was blotted off and the tissue was minced, then homogenized in TRI reagent (MRC). The samples were centrifuged in the presence of bromochloropropane and sodium acetate. The RNA-containing upper layer was removed and the RNA purified on an RNeasy extraction column (Qiagen). The sample was treated with an on-column DNase treatment (RNase-free DNase, Qiagen). The purity and quantity of total RNA was evaluated with the RNA 6000 NanoLabChip in an Agilent Bioanalyzer.
|
| Label |
Alexa-647
|
| Label protocol |
Biotinylated cRNA was prepared using the MessageAmp II-Biotin Enhanced Kit from Ambion from 2 micrograms total RNA starting material.
|
| |
|
| Hybridization protocol |
10 micrograms of biotinylated cRNA were mixed with fragmentation buffer and heated to 94C for 20 min. The fragmented cRNA was mixed with CodeLink hybridization buffer, loaded on the microarray slides, and hybridized for 18 hours at 37C.
|
| Scan protocol |
The slides were scanned in an Axon GenePix 4000B scanner.
|
| Description |
Protocol for dissecting the mesenteric arterial tree is detailed in Rodrigo MC et al., J Pharmacol Toxicol Methods 2002;47(2):87-92.
|
| Data processing |
global median normalization, CodeLink Expression Analysis 4.1.0.4163
|
| |
|
| Submission date |
Jul 19, 2010 |
| Last update date |
Jul 06, 2011 |
| Contact name |
Kathleen M Eyster |
| E-mail(s) |
Kathleen.Eyster@usd.edu
|
| Organization name |
University of South Dakota
|
| Department |
Basic Biomedical Sciences
|
| Street address |
414 E. Clark St.
|
| City |
Vermillion |
| State/province |
SD |
| ZIP/Postal code |
57069 |
| Country |
USA |
| |
|
| Platform ID |
GPL2896 |
| Series (1) |
| GSE23037 |
Estrogens and selective estrogen receptor modulators regulate gene and protein expression in the mesenteric arteries |
|
