|
| Status |
Public on Jul 24, 2017 |
| Title |
OVSAHO MI-res2 |
| Sample type |
RNA |
| |
|
| Source name |
Epithelial Ovarian Carcinoma cells treated with platinum
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: ovarian cell line
histotype: serous
|
| Treatment protocol |
not applicable
|
| Growth protocol |
not applicable
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from MCL cells using the TRIZOL Reagent (ThermoFisher) and validated for integrity and purity using the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.1 ug RNA using the One-Color Low Input Quick Amp Labeling Kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
1.6 ug of Cy3-labelled cRNA (specific activity >6.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 55 ul containing 25x Agilent fragmentation buffer and 10x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 55 ul of 2x Agilent Hi-RPMI hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Whole Human Genome Oligo Microarrays (G4112A) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then scan immediately.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505B) using one color scan setting for 1x44k array slides (Scan Area 61x21.6 mm, Scan resolution 10um, Dye channel is set to Green and Dynamic Range of Green PMT is set to 100% and 10%).
|
| Description |
Constitutive Gene expression of EOC cells.
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.7.3.1 (Agilent) using default parameters (protocol GE1-v107_Sep09 and Grid: 014850_D_F_20100430) to obtain background subtracted and spatially detrended Processed Signal intensities.
|
| |
|
| Submission date |
Jan 18, 2017 |
| Last update date |
Jul 24, 2017 |
| Contact name |
Riccardo Bomben |
| E-mail(s) |
rbomben@cro.it
|
| Phone |
+39 0434 659718
|
| Organization name |
CRO Aviano
|
| Department |
D-RT
|
| Lab |
Clinical and Experimental Onco Hematology Unit
|
| Street address |
Via Franco Gallini 2
|
| City |
Aviano |
| ZIP/Postal code |
33080 |
| Country |
Italy |
| |
|
| Platform ID |
GPL17077 |
| Series (2) |
| GSE93793 |
Characterization of three Epithelial Ovarian Cancer cell lines resistant to cisplatin (mRNA). |
| GSE93795 |
Characterization of three Epithelial Ovarian Cancer cell lines resistant to cisplatin. |
|
