|
| Status |
Public on Dec 31, 2011 |
| Title |
Day0 b VSN |
| Sample type |
RNA |
| |
|
| Source name |
undifferentiated murine embryonic stem cell
|
| Organism |
Mus musculus |
| Characteristics |
cell line: ES-D3 murine pluripotent cells
atcc catalog#: CRL-1934
time: day 0
|
| Biomaterial provider |
Axiogenesis AG Germany
|
| Treatment protocol |
ES cells were trypsinized at d-2, seeded onto a gelatin-coated 6cm TC dish (Becton Dickinson, Heidelberg, Germany), and cultured for 48h in DMEM 15% FCS. The timing of this last passage was in parallel with the ES cell passage used to initiate differentiation.
|
| Growth protocol |
aPIG 44 cells were cultured on mouse embroynic fibroblasts in high-glucose Dulbecco’s modified Eagle’s medium (DMEM) supplemented with nonessential amino acids (0.1 mM), L-glutamine (2 mM), beta-mercaptoethanol (0.1 mM), LIF (ESGR) (500 U/ml), neomycin (6 µg/mL), and batch-tested fetal calf serum (FCS) (15% v/v) (all Invitrogen, Karlsruhe, Germany).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNAs were extracted by using PeqGold RNApure (Peqlab Biotechnology, Erlangen, Germany) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Total RNA containing low molecular weight RNA was labelled using the flashtag RNA labeling kit (Genisphere, Hatfield, PA, USA) according to the manufacturer’s instructions.
|
| |
|
| Hybridization protocol |
Sample was hybridized to a GeneChip® miRNA Array (Affymetrix, Santa Claraq, CA, USA) at 48°C and 60 rpm for 16 hours then washed and stained on Fluidics Station 450 (Fluidics script FS450_0003).
|
| Scan protocol |
Microarray was finally scanned on a GeneChip® Scanner 3000 7G (Affymetrix).
|
| Description |
mouse ES cells (D3, ATCC CRL 1934) transfected with the alpha-MHC-Pac-IRES-EGFP vector
|
| Data processing |
Data was normalized with VSN (Bioconductor, vsn package).
Normalization and probe set summarization was carried out on the mouse-specific probe sets only.
|
| |
|
| Submission date |
Sep 10, 2010 |
| Last update date |
Dec 31, 2011 |
| Contact name |
Lin Gan |
| Organization name |
University Hospital Aachen
|
| Department |
IZKF
|
| Lab |
Genomics facility
|
| Street address |
Pauwelsstr. 30
|
| City |
Aachen |
| ZIP/Postal code |
52074 |
| Country |
Germany |
| |
|
| Platform ID |
GPL8786 |
| Series (1) |
| GSE24066 |
miRNA profiling during cardiomyocyte-specific differentiation of murine embryonic stem cells based on two different miRNA array platforms |
|
