|
| Status |
Public on Jan 01, 2010 |
| Title |
LTED_Replicate 1 |
| Sample type |
RNA |
| |
|
| Source name |
LTED
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: MCF-7
tissue type: breast cancer
|
| Treatment protocol |
Cells were grown constantly in the presence of testosterone (where indicated) and inhibitor.
|
| Growth protocol |
Cells were grown long-term in the absence and presence of testosterone (T). Where indicated, testosterone is labeled at T+inhibitor. Cells were grown in charcoal-dextran stripped FBS containing MEM.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol Extraction for total RNA isolation, according to the maufacturer's recommendation.
|
| Label |
Cy3
|
| Label protocol |
100ng total RNA was used for labeling reaction with pCp-Cy3 for 2 hours. Following labeling protocol, labeled RNA was purified with micro Bio-Spin 6 columns (Bio-Rad 732-6221), according to manufacturers recommendations.
|
| |
|
| Hybridization protocol |
Labeled RNA samples were dried using a speed vac and resuspended in Dnase/Rnase-free water. Total amount of labeled RNA was mixed with blocking agent and hybridization buffer (provided with Agilent miRNA Labeling Reagent and Hybridization kit 5190-0408). Hybridization was carried out at 55degreesC for 20hours in the Agilent hybridization chamber G2534A. After hybridization chips were washed with the Gene expression wash buffer kit (Agilent 5188-5327) twice for 5 minutes.
|
| Scan protocol |
Slides were scanned using the Agilent scanner G2565BA for the one color setting. Scan area for the 8x15K format was set at 61x21.6 mm, with a scan resolution of 5um, green dye channel and Green PMT set to 100%.
|
| Description |
microRNA expression before and after resistance
|
| Data processing |
Scanned data was analyzed with Feature Extraction (FE) software version 9.5.1.
|
| |
|
| Submission date |
Aug 24, 2009 |
| Last update date |
Aug 24, 2009 |
| Contact name |
Selma Masri |
| E-mail(s) |
[email protected]
|
| Organization name |
City of Hope
|
| Department |
Tumor Cell Biology
|
| Lab |
Shiuan Chen Lab
|
| Street address |
1500 E Duarte Rd
|
| City |
Duarte |
| State/province |
CA |
| ZIP/Postal code |
91010 |
| Country |
USA |
| |
|
| Platform ID |
GPL9081 |
| Series (1) |
| GSE17775 |
microRNA expression profiles in Aromatase Inhibitor-Resistant, Tamoxifen-Resistant and LTED breast cancer cell lines. |
|
