|
| Status |
Public on Oct 28, 2006 |
| Title |
A2780_6h_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
A2780 ovarian carcinoma cell line
|
| Organism |
Homo sapiens |
| Characteristics |
A2780 ovarian carcinoma cell line
|
| Biomaterial provider |
ECACC (Cat no.93112519)
|
| Treatment protocol |
Human Ovarian cell line A2780 was untreated or exposed to 3 micromolar of a cell cycle inhibitor for 6hrs
|
| Growth protocol |
A2780 was grown in RMPI 1640 + 10% FCS +2mM Glutamine and maintained at 37C in an atmosphere of 5% CO2 and 95% room air. Cells were plated at 2 x 106 cell/ml and after 24hrs were exposed to 3 micromolar of the inhibitor for 6hrs.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extraction was performed using Qiagen RNeasy Mini kit 74106 following manufacturers instructions.
|
| Label |
biotin
|
| Label protocol |
5 micrograms of each RNA samples was used for the amplification/labelling reaction. 1st and 2nd strand cDNA synthesis was performed with the Invitrogen kit (11917-020) and the IVT reaction was done with Megascript T7 from Ambion (1334). All steps were done according to manufacturers instructions and cRNA was quantitated on a spectrophotometer. 15 micrograms of cRNA was fragmented and checked by denaturing gel electrophoresis. Bacterial transcripts at the cRNA level were spiked into each sample prior to hybridisation.
|
| |
|
| Hybridization protocol |
10 micrograms of fragmented cRNA was hybridised to a Human Genome U133 Plus 2.0 Array (900466). Hybridisations, washing and staining were performed according to Affymetrix protocols
|
| Scan protocol |
Hybridised arrays were scanned with the Genechip Scanner 3000
|
| Description |
Human Ovarian cell line A2780 was obtained from ECACC (Cat no.93112519) and cells were untreated or exposed to 3 micromolar of a cell cycle inhibitor for 6hrs. Three biological replicates were performed for each treatment. Only attached cells were harvested. RNA was purified using a Qiagen RNA purification kit. RNA was quantitated using a spectrophotometer and the quality of the RNA was assessed using a Bioanalyser.
Biological replicates were pooled to obtain a unique sample for each treatment, which was then divided to generate three aliquots for each condition (technical replicates).
|
| Data processing |
Probe set intensities were calculated using the RMA algorithm and normalized by the quantiles method
|
| |
|
| Submission date |
Oct 26, 2006 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Roberta Bosotti |
| E-mail(s) |
roberta.bosotti@nervianoms.com
|
| Organization name |
Nerviano Medical Sciences srl
|
| Department |
Biotechnology
|
| Lab |
Genomics
|
| Street address |
viale Pasteur,10
|
| City |
Nerviano |
| State/province |
MI |
| ZIP/Postal code |
20014 |
| Country |
Italy |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE6140 |
Cross platform microarray analysis for robust identification of differentially expressed genes |
|
| Relations |
| Reanalyzed by |
GSE64985 |
| Reanalyzed by |
GSE119087 |
