|
| Status |
Public on Sep 29, 2007 |
| Title |
Luc RNAi biological rep5 |
| Sample type |
RNA |
| |
|
| Source name |
E14 mouse embryonic stem cells
|
| Organism |
Mus musculus |
| Characteristics |
Luc shRNA (targeting Firefly luciferase transcript) transfected E14 mouse embryonic stem cells
|
| Treatment protocol |
2 ug of Luc shRNA (targeting Firefly luciferase transcript) was cloned in pSUPER puro vector (OligoEngine, Seattle, WA) and transfected into E14 mouse embryonic stem cells cells on 60mm plates. Puromycin (Sigma) selection was introduced 1 day after transfection at 1.0 µg/ml, and maintained for 4 days prior to harvesting RNA
|
| Growth protocol |
Feeder-free E14 mouse embryonic stem cells were cultured at 37°C with 5% CO2. All cells were maintained on gelatin-coated dishes in Dulbecco’s modified Eagle medium (DMEM; GIBCO), supplemented with 15% heat-inactivated fetal bovine serum (FBS; GIBCO), 0.055 mM Beta-mercaptoethanol (GIBCO), 2 mM L-glutamine, 0.1 mM MEM nonessential amino acid, 5,000 units/ml penicillin/streptomycin and 1,000 units/ml of LIF (Chemicon)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using Trizol (Invitrogen) and purified with RNeasy minikit (Qiagen).
|
| Label |
Biotin
|
| Label protocol |
Biotinylated cRNA were prepared from 500 ng of total RNA according to the standard protocol (TotalPrep RNA Amplification Kit, Instruction Manual, Catalog #IL1791, Illumina).
|
| |
|
| Hybridization protocol |
1.5 ug of cRNA were hybridized to Illumina Mouse-6 Expression BeadChip v1 for 20 hr at 55oC on Ilumina Hybridization Oven.
|
| Scan protocol |
Illumina Mouse-6 Expression BeadChip v1 were washed and stained with streptavidin-Cy3 before scanning using the Illumina Bead Reader.
|
| Description |
Gene expression data from Luc shRNA treated E14 embryonic stem cells
|
| Data processing |
The data were analyzed using Bead Array suite software. Rank invariant normalization method was chosen to obtain the signal intensity.
|
| |
|
| Submission date |
Sep 01, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
Huck Hui Ng |
| E-mail(s) |
[email protected]
|
| Phone |
+65 478 8000
|
| URL |
http://tinyurl.com/39ut7w
|
| Organization name |
Genone Institute of Singapore
|
| Department |
Stem Cell and Development biology
|
| Street address |
Biopolis street
|
| City |
Singapore |
| State/province |
Singapore |
| ZIP/Postal code |
137688 |
| Country |
Singapore |
| |
|
| Platform ID |
GPL4234 |
| Series (1) |
| GSE8937 |
Jmjd1a / Jmjd2c RNAi expression profile |
|
