|
| Status |
Public on Feb 03, 2009 |
| Title |
Tamoxifen 3 |
| Sample type |
RNA |
| |
|
| Source name |
postmenopausal endometrium, Tamoxifen 3
|
| Organism |
Macaca fascicularis |
| Characteristics |
Adult female ovariectomized macaque
normal endometrium
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from frozen endometrial samples using Tri Reagent (Molecular Research Center, Cincinnati,OH); purified using RNeasy Mini Kit (Qiagen, Valencia,CA); and quantified using NanoDrop ND1000 UV-vis spectrophotometer(NanoDrop, Wilmington,DE). RNA intactness and quality was confirmed using an Agilent 2100 Bioanalyzer (Wilmington,DE).
|
| Label |
Biotin dye
|
| Label protocol |
Enzo Bioarray kit. Biotinylated cRNA samples were prepared according to the standard Enzo BioarrayTM protocol (Enzo Life Sciences). In brief, one ug of total RNA from each sample was converted to double-stranded cDNA with the BioarrayTM Single-Round RNA Amplification and Labeling Kit (Enzo Life Sciences). After second-strand synthesis, the cDNA was purified with the cDNA Purification Kit (Enzo Life Sciences). The resulting double-stranded DNA was then used to generate multiple copies of biotinylated cRNA by in vitro transcription with the BioarrayTM HighYieldTM RNA Transcript Labeling Kit (Enzo Life Sciences).
|
| |
|
| Hybridization protocol |
Standard Affymetrix protocol for Eukaryotic samples. For each sample, ten µg of biotinylated cRNA spiked with bioB, bioC, bioD and cre (Hybridization Control) was hybridized to a Rhesus Macaque Microarray for 16 hours at 45ºC. Following hybridization, all arrays were washed and stained in an Affymetrix GeneChip Fluidics Station.
|
| Scan protocol |
Stained arrays were scanned with an Affymetrix GeneChip® Scanner 3000.
|
| Description |
Data were extracted from scanned images using GeneChip Operating Software(Affymetrix, Inc., Santa Clara,CA.
|
| Data processing |
Quality checks and data analyses were carried out using Affymetrix GeneChip Operating Software (GCOS) and Expression Console. The samples were analyzed using the MAS5 algorithm in Affymetrix Expression Console.
|
| |
|
| Submission date |
Jan 22, 2009 |
| Last update date |
Feb 02, 2009 |
| Contact name |
Charles Wood |
| E-mail(s) |
chwood@wfubmc.edu
|
| Organization name |
Wake Forest University School of Medicine
|
| Department |
Pathology
|
| Street address |
Medical Center Blvd
|
| City |
Winston-Salem |
| State/province |
NC |
| ZIP/Postal code |
27157-1040 |
| Country |
USA |
| |
|
| Platform ID |
GPL3535 |
| Series (1) |
| GSE14518 |
Endometrial profile of low-dose estradiol and tamoxifen combination therapy |
|
