|
| Status |
Public on Jul 22, 2009 |
| Title |
MCF-7 E2 4h_1 |
| Sample type |
RNA |
| |
|
| Source name |
MCF-7 cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: MCF-7 cells
agent: E2 (100 nM) treatment
time point: 4 hr
|
| Treatment protocol |
MCF-7 cells cultured in phenol red-free DMEM containing 10% charcoal-stripped FBS for 2 days and E2 (100 nM) treatment for 4 hr.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
To isolate miRNA fractions from total RNA, 100μｇ total RNA was fractionated and cleaned up with the flashPAGETM fractionator System and reagents (Ambion) per the manufacturer’s recommendation.
|
| Label |
Cy5
|
| Label protocol |
mirVanaTM miRNA Labeling Kit (Ambion) and amine-reactive dyes as recommended by the manufacturer.
|
| |
|
| Hybridization protocol |
A 3x miRNA Hybridization Buffer (Ambion) was added to the fluorescently labeled miRNAs and the solution was heated at 95ºC for 3 min. Slides were hybridized form 12 to 16 h at 42 ºC in sealed cassettes using a water bath. Following hybridization, the slides were washed and dried prior to a high-resolution scan.
|
| Scan protocol |
Scan on a GenePix 4000B (Axon Instruments, CA, USA). Raw data were normalized and analyzed using Array-Pro Analyzer Version 4.5 (Media Cybernetics, Inc., Silver Spring, MD, USA).
|
| Description |
E2 treatment for 4 hr of MCF-7 cells
|
| Data processing |
We used Array-Pro Analyzer Ver4.5 (Media Cybernetics, Inc., Silver Spring, MD, USA) to determine the signal intensity of each spot and its local background on microarrays. We calculated net intensity by subtracting the mean intensity of all pixels within the local background area from the mean intensity of all pixels within the spot areas.
|
| |
|
| Submission date |
Jun 21, 2009 |
| Last update date |
Jul 22, 2009 |
| Contact name |
kaoru yamagata |
| E-mail(s) |
ykaoru7@hotmail.com
|
| Organization name |
university of tokyo
|
| Street address |
yayoi 1-1-1
|
| City |
bukyo-ku |
| State/province |
Tokyo |
| ZIP/Postal code |
113-0032 |
| Country |
Japan |
| |
|
| Platform ID |
GPL8700 |
| Series (1) |
| GSE16737 |
Global survey of miRNA microarray of MCF-7 cells with or without estrogen (E2) treatment |
|
