|
| Status |
Public on Sep 01, 2013 |
| Title |
HCT-15 second |
| Sample type |
RNA |
| |
|
| Source name |
colorectal cancer cell line
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HCT-15
|
| Growth protocol |
RPMI 1640, Leibovitz's L-15 Medium, McCoy's 5A, or D-MEM/F-12, containing 10% heat-inactivated fetal bovine serum and 1% penicillin-streptomycin. The cell lines were cultured at 37°C in a humidified atmosphere of 5% CO2 and 95% air in a plastic flask.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Samples were extracted using Qiagen Rneasy kit.
|
| Label |
biotin
|
| Label protocol |
From 2 µg of total RNA, double-stranded cDNA and subsequent biotinlabeled cRNA were synthesized using the CodeLink Expression Assay Kit according to the manufacturer’s instructions.
|
| |
|
| Hybridization protocol |
10 µg of fragmented cRNA was applied to each bioarray. Bioarrays were hybridized overnight at 37 ºC at 300 rpm in an incubator, then washed in stringent TNT buffer at 46 ºC for 1 h followed by incubation with streptavidin-Cy5 at room temperature for 30 min in the dark. Bioarrays were washed in TNT buffer twice each followed by a rinse in 0.05% Tween-20 in water and then dried.
|
| Scan protocol |
Glass slides were scanned using a GenePix 4000A scanner.
|
| Data processing |
The grids of the image spots were adjusted and their signals were analyzed using the CodeLink System Software, version 5.0, which produces both a raw and a normalized hybridization signal for each spot on the array.
|
| |
|
| Submission date |
Aug 12, 2013 |
| Last update date |
Sep 01, 2013 |
| Contact name |
Yasufumi Matsuda |
| E-mail(s) |
y-matsuda@surg1.med.tohoku.ac.jp
|
| Phone |
81-22-717-7205
|
| Organization name |
Tohoku University Graduate School of Medicine
|
| Department |
Surgery
|
| Lab |
Biological-Regulation and Oncology
|
| Street address |
1-1 seiryouchou aobaku
|
| City |
Sendai |
| State/province |
Miyagi |
| ZIP/Postal code |
980-8574 |
| Country |
Japan |
| |
|
| Platform ID |
GPL2891 |
| Series (1) |
| GSE49811 |
Expression profiles of colorectal cancer cell lines |
|
