|
| Status |
Public on Aug 06, 2016 |
| Title |
MCF10A control |
| Sample type |
RNA |
| |
|
| Source name |
MCF10A control
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: Human mammary epithelial cell (MCF10A)
agent: control
|
| Treatment protocol |
MCF10A cells were purchased from American Type Culture Collection. The MCF10A were routinely cultured in mammary epithelial cell growth medium kit at 37ºC in 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted by using the RNeasy mini kit (Qiagen, Tokyo, Japan).
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.1ug Total RNA using the Low Input Quick Amp Labeling Kit(Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
0.6 ug of Cy3-labelled cRNA was fragmented at 60°C for 30 minutes in a reaction volume of 25 ul containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 25 ul of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Microarrays for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2565CA) using one color scan setting for 8x60k array slides (Scan Area 61x21.6 mm, Scan resolution 3um, Dye channel is set to Green PMT is set to 100%).
|
| Description |
Gene expression control
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.10 (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities.
|
| |
|
| Submission date |
Aug 05, 2016 |
| Last update date |
Aug 06, 2016 |
| Contact name |
Naoki Nanashima |
| E-mail(s) |
n_nanashima@auhw.ac.jp
|
| Organization name |
Aomori University of Health and Welfare
|
| Department |
Department of Nutrition
|
| Street address |
58-1 Mase, Hamadate
|
| City |
Aomori |
| State/province |
Aomori |
| ZIP/Postal code |
030-8505 |
| Country |
Japan |
| |
|
| Platform ID |
GPL17077 |
| Series (1) |
| GSE85235 |
Effects of black currant extract on gene expression on MCF10A |
|
