|
| Status |
Public on Sep 29, 2011 |
| Title |
ME:MDA_MB_435 [118180hp133a11] |
| Sample type |
RNA |
| |
|
| Source name |
Ductal carcinoma-mammary gland; breast; duct; metastatic site: pleural effusion;
|
| Organism |
Homo sapiens |
| Characteristics |
tissue of origin: Melanoma
cell line: MDA_MB_435
age: 31
Sex: F
epithelial: no
source: Pleural effusion
ploidy: 2n+, Hyperdiploid (47-57)
p53 mutation: MT
doubling time: 25.8
contributing person: R. Cailleau
reference: Cancer Res 40:3118-3129,1980
|
| Treatment protocol |
No treatment
|
| Growth protocol |
Cells grown in RPMI 1640 with 5% FBS (Bio Whittaker, not heat inactivated) and 1% L-glutamine. Cells not grown past 80% confluencey for attached cells, or 0,5 x 10-6 cells for suspended. Trypsinize (for attached cells) cells with 5 ml trypsin-EDTA per T162, for 15 min., at 370C. Spin down suspended cells, and resuspend in ~3ml growth media. Count cells. Pass 1x10-6 cells into new T162’s w 30 ml media. Repeat growth cycle until desired amount of cells are available. Cells are not grown past passage 20.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Growth schedule prior to harvest: Grow cells to ~80 confluencey. Re-feed cells the day prior to harvest. Draw off media from attached cells, or spin down suspended cells. Rinse cells with ~10 ml 1x PBS. Purify using Quiagen Midi Kit (Cat # 75144) per instructions. In brief, lyse cells in 15 ml lysis buffer (w 10 ul fresh BME per ml) per 4 T162s. Scrape (attached) cells. Pipet into a 50 ml tube. Repeat with the next set of 4 flasks. When done, vortex 10 sec.. Draw lysate thru a 20 guage needle 12x’s. Freeze lysate at -80 deg C. Use a maximum of 100 x 10-6 cells per column.
|
| Label |
biotin
|
| Label protocol |
Done by Genelogic according to manufacturer's recommendations
|
| |
|
| Hybridization protocol |
Done by Genelogic according to manufacturer's recommendations
|
| Scan protocol |
Affymetrix MicroArraySuite
|
| Description |
Ductal carcinoma-mammary gland; breast; duct; metastatic site: pleural effusion;
|
| Data processing |
Probes were normalized using RMA in Partek Genomics Suite version 6.2
|
| |
|
| Submission date |
Sep 29, 2011 |
| Last update date |
Sep 29, 2011 |
| Contact name |
Sudhir Varma |
| E-mail(s) |
sudhirv4rma@gmail.com
|
| Organization name |
HiThru Analytics
|
| Street address |
1215 Wessex Pl
|
| City |
Princeton |
| State/province |
NJ |
| ZIP/Postal code |
08540 |
| Country |
USA |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE32474 |
Comparison between cell lines from 9 different cancer tissue (NCI-60) (Affymetrix U133 Plus 2.0) |
|
