|
| Status |
Public on May 01, 2013 |
| Title |
ESCs_AllStars_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
VV3 ESCs, control siRNA transfected
|
| Organism |
Mus musculus |
| Characteristics |
strain/background: 129SV
gender: male
cell line: VV3
cell type: embryonic stem cells (ESCs)
sirna: non-silencing negative control
|
| Treatment protocol |
ESCs were transfected with non-silencing negative control siRNA (AllStars) using Lipofectamine RNAiMAX. After 3 days culture, the live and single cells were purified using FACS from the ESCs.
|
| Growth protocol |
Cells were cultured with ESCs medium with 14% KSR, 1% FBS and LIF at 37 ºC in a humidified incubator with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was purified using the RNeasy micro kit according to the manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
Cy3-labeled cRNA samples were prepared using the Agilent Low Input Quick Amp Labeling Kit (one-color) (5190-2305) and RNA Spike-In Kit (one-color) (5188-5282) according to the manufacturer's instructions. Input was 100 ng of total RNA from each sample.
|
| |
|
| Hybridization protocol |
1.65ug of Cy3-labelled cRNAs from each sample were fragmented and hybridized to microarrays according to the manufacturer's instructions using the Gene Expression Hybridization kit (Agilent 5188-5242) and Agilent hybridization oven. After hybridization, microarrays were washed using the Gene Expression Wash Pack (Agilent 5188-5327).
|
| Scan protocol |
The microarrays were scanned using the Agilent DNA microarray scanner (Agilent G2505C); the spot signals were quantified using Feature Extraction software.
|
| Description |
Gene expression of negative control siRNA-transfected VV3 mouse ESCs.
|
| Data processing |
Data was normalized and analyzed by GeneSpring GX (Agilent).
|
| |
|
| Submission date |
Mar 14, 2013 |
| Last update date |
May 01, 2013 |
| Contact name |
Ikuma Maeda |
| E-mail(s) |
[email protected]
|
| Phone |
+81-22-717-8572
|
| Fax |
+81-22-717-8573
|
| Organization name |
Institute of Development, Aging and Cancer, Tohoku University
|
| Lab |
Cell Resource Center for Biomedical Research
|
| Street address |
4-1 Seiryomachi, Aoba-ku
|
| City |
Sendai |
| State/province |
Miyagi |
| ZIP/Postal code |
980-8575 |
| Country |
Japan |
| |
|
| Platform ID |
GPL11202 |
| Series (1) |
| GSE45181 |
Max is a repressor of germ-cell-related gene expression in mouse embryonic stem cells |
|
