|
| Status |
Public on Sep 30, 2012 |
| Title |
HTB58_controlsiRNA_2 |
| Sample type |
RNA |
| |
|
| Source name |
Adherent lung cancer cell line, control siRNA
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HTB-58
gender: male
age: 65 years
race: Caucasian
disease: squamous cell carcinoma of the lung
sirna: control
|
| Treatment protocol |
Cell lines were transfected with 1 μg/μl (75 μM) of chemically synthesized anti-EVI1 siRNA or control siRNA.
|
| Growth protocol |
Cells were cultured in RPMI-1640 medium supplemented with 10% FBS in the case of HTB-58 cells or with 20% FBS and 5-10 ng/ml recombinant GM-CSF in the case of TF-1.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The cells were harvested with TRIzol Reagent (Invitrogen) and the RNA was extracted according to the manufacturer's instructions. As a last step of the extraction procedure, the RNA was purified with the RNeasy Mini-kit (Qiagen, Hilden, Germany).
|
| Label |
biotin
|
| Label protocol |
The cDNA was prepared from 2 microg total RNA using the One –Cycle cDNA Synthesis kit from Affymetrix. This cDNA was used for synthesizing the cRNA using the IVT labeling kit (Affymetrix). The synthesized cRNA was purified with the GeneChip Sample Cleanup Module (Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 microg of cRNA were hybridized to the Affymetrix HG-U133Plus2 chip according to the protocols provided by the manufacturer.
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000 from Affymetrix.
|
| Description |
Replicate 2 of 3.
|
| Data processing |
RMA normalization.
|
| |
|
| Submission date |
May 26, 2009 |
| Last update date |
Sep 30, 2012 |
| Contact name |
Maria D Odero |
| E-mail(s) |
modero@unav.es
|
| Organization name |
University of Navarra
|
| Street address |
Pio XII, 55
|
| City |
Pamplona |
| ZIP/Postal code |
31008 |
| Country |
Spain |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE16238 |
Genome-wide identification of binding sites of Evi1 in human myeloid cell lines |
|
