|
| Status |
Public on Jan 28, 2010 |
| Title |
hsa-mir-182 inhibitor_17-3 |
| Sample type |
RNA |
| |
|
| Source name |
ES2 ovarian cancer cell line
|
| Organism |
Homo sapiens |
| Characteristics |
protocol: hsa-mir-182 depletion
|
| Treatment protocol |
Transfection conditions for each clear cell ovarian cancer cell line were optimized for maximum efficiency in preliminary experiments using Dy547-labeled miRIDIAN hairpin inhibitor and mimic transfection controls (Dharmacon). ES-2 cells were transfected in 6-well plates (2.5 x 105 cells/well) using 10 μl X-tremeGENE siRNA Transfection Reagent (Roche) and 2 μg hsa-mir-182 hairpin inhibitor (50 nM final concentration) (Dharmacon). Control groups were cells treated with transfection reagent alone (mock transfection), and cells transfected with miRIDIAN hairpin inhibitor negative control #1 or mimic negative control #2, which are based on C. elegans miRNAs that have minimal sequence identity to human miRNAs (Dharmacon).
|
| Growth protocol |
ES-2 cells were cultured in DMEM/F12 (Invitrogen), with 10% heat-inactivated fetal bovine serum and 1% penicillin-streptomycin solution (Invitrogen).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were harvested 48 hours after transfection. Total RNA was extracted using the mirVana miRNA Isolation Kit (ABI). RNA quality were inspected on a 2100 Bioanalyzer (Agilent).
|
| Label |
biotin
|
| Label protocol |
According to Illumina protocol.
|
| |
|
| Hybridization protocol |
According to Illumina protocol.
|
| Scan protocol |
According to Illumina protocol.
|
| Description |
hsa-mir-182 inhibitor (mir-182 knockdown in ES2), biological rep3
|
| Data processing |
The image data were compiled into a project using Beadstudio software gene expression module version 3.2.7. No normalization was used. The data was exported as txt files using the build-in function of export to GeneSpring GX format. The option of sample probe profile was chosen. Data were subsequently quantile normalized for the analysis.
|
| |
|
| Submission date |
Jun 12, 2009 |
| Last update date |
Jan 28, 2010 |
| Contact name |
Chad Creighton |
| E-mail(s) |
creighto@bcm.tmc.edu
|
| Organization name |
Baylor College of Medicine
|
| Department |
Biostatistics, Ducan Cancer Center
|
| Street address |
One Baylor Plaza, Mail Stop: BCM305
|
| City |
Houston |
| State/province |
TX |
| ZIP/Postal code |
77030 |
| Country |
USA |
| |
|
| Platform ID |
GPL6947 |
| Series (2) |
| GSE16572 |
Gene expression analyses of mir-182 knockdown in ovarian clear cell cancer cell line |
| GSE16574 |
Gene expression analyses of putative miRNA targets in ovarian clear cell cancer cell lines |
|
