|
| Status |
Public on Mar 11, 2014 |
| Title |
Iliac artery necropsy_Equol treatment_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Iliac artery necropsy_high fat diet_Equol treatment
|
| Organism |
Macaca fascicularis |
| Characteristics |
strain: Cynomolgus monkey
age: greater than 11 years
gender: female
high fat diet: 6.5 years
ovariectomized: ~4 years
tissue: iliac artery necropsy
treatment: Equol
duration: 8 months
atherosclerotic plaque size pretreatment: 1.003 mm2
atherosclerotic plaque size postreatment: 0.951 mm2
animal id: 5530
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The iliac artery samples were obtained at necropsy. The artery samples were placed in RNAlater (Ambion) in the surgical suite. For processing, RNAlater was blotted off and the artery sample was minced with scalpels, and homogenized in TRI reagent (MRC) with a Polytron (Brinkmann). 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 0.275 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 |
Approval for all animal procedures was obtained from the Institutional Animal Care and Use Committee.
|
| Data processing |
global median normalization, CodeLink Expression Analysis 4.1.0.4163
|
| |
|
| Submission date |
Apr 06, 2012 |
| Last update date |
Mar 11, 2014 |
| 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 |
GPL2895 |
| Series (1) |
| GSE37186 |
Effects of equol on gene expression in female cynomolgus monkey iliac arteries |
|
