|
| Status |
Public on Nov 11, 2011 |
| Title |
B16-F10 cells exposed to 100 μM CoCl2, biological replica 2, experiment 3 |
| Sample type |
RNA |
| |
|
| Source name |
B16-F10 cells exposed to hypoxia mimicry
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6J
cell line: B16-F10
cell origin: melanoma
biomaterial provider: ATCC CRL-6475T
condition: exposed to 100 μM CoCl2
|
| Treatment protocol |
Hypoxic cpnditions were introduced by incubating the cells in 1% oxygen using either Billubs-Rothenberg chamber or oxygen flow-regulated incubator for 24 h. Hypoxia-mimicry was performed by treating the cells with either 100 μM or 200 μM CoCl2 for 24 h.
|
| Growth protocol |
The mouse melanoma B16-F10 cells were grown in CO2 incubator at 37°C in RPMI medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum (ICN).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using Rneasy Mini Kit (Qiagen) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Total RNA (8 μg) was used for synthesis of double stranded cDNA according to manufacturer's instructions. Half of the cDNA volume was used for synthesis of biotynylated cRNA with the BioArray High Yield RNA Transcript Labeling Kit (Enzo Diagnostics).
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| |
|
| Hybridization protocol |
Following fragmentation, 16 μg of cRNA were hybridized for 16 hr at 45ºC on Mouse Expression Arrays 430A (Affymetrix). GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
Arrays were scanned using Hewlett-Packard GeneArray G2500A scanner with MAS5 software (Affymetrix).
|
| Description |
Gene expression data from B16-F10 cells exposed to 100 μM CoCl2 for 24 hours
|
| Data processing |
Data was acquired using GCOS 1.2 software (Affymetrix). The preprocessing was performed by Robust Multiarray Analysis (RMA, Bioconductor).
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| |
|
| Submission date |
Nov 10, 2011 |
| Last update date |
Nov 11, 2011 |
| Contact name |
Michal Swierniak |
| Organization name |
University of Warsaw
|
| Department |
Centre of New Technologies
|
| Street address |
Banacha 2C
|
| City |
Warsaw |
| ZIP/Postal code |
02-097 |
| Country |
Poland |
| |
|
| Platform ID |
GPL81 |
| Series (1) |
| GSE33607 |
Expression data from mouse B16-F10 cells exposed to hypoxic conditions in vitro. |
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