|
| Status |
Public on Feb 03, 2014 |
| Title |
Control EBs on day 3 A |
| Sample type |
RNA |
| |
|
| Source name |
No doxycycline treated iEGS, ES cell
|
| Organism |
Mus musculus |
| Characteristics |
tissue: embryonic stem cell
genotype: wild-type
timepoint: D3
treatment: -DOX
replicate: 1
|
| Treatment protocol |
Doxycycline (1µg/ml) was added in iEGS+DOX group from day 2 to 3.
|
| Growth protocol |
Mouse ES cells were maintained on PMEF cells in DMEM with 15% pre-selected fetal calf serum (FCS), 2% LIF, 1% L-glutamine, 1% NEAA and 4.5x 10–4 M MTG. ES cells were differentiated in IMDM containing 15% differentiation-screened FCS, 1% L-glutamine, 50 g/ml ascorbic acidand 4.5x 10–4 M MTG.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA were extracted and purified by RNeasy Mini Kit (Qiagen.) cDNA synthesis and amplification was performed using the WT cDNA synthesis and amplification kit (Affymetrix).
|
| Label |
Biotin
|
| Label protocol |
Biotinylated cDNA were prepared according to the standard NuGen WT-Ovation Pico RNA Amplification system from 500pg-50 ng total RNA.
|
| |
|
| Hybridization protocol |
Following fragmentation with NuGen Encore Biotin Module, 2.5 ug of cDNA were hybridized on Exon 1.0 ST Array in the GeneChip Hybridization Oven 640 for 18 hr at 45C. GeneChip were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 7G 3000.
|
| Description |
Control EBs on day 3
|
| Data processing |
Data were processed using Affymetrix Expression Console software. Gene level expression were summarized from the intensities and then normalized using the Robust Multiarray Analysis (RMA) algorithm.
probe_group_file: MoGene-1_1-st-v1.r4.pgf
meta-probeset file: MoGene-1_1-st-v1.r4.mps
|
| |
|
| Submission date |
Mar 13, 2013 |
| Last update date |
Feb 03, 2014 |
| Contact name |
Fang Liu |
| E-mail(s) |
fliu@pathology.wustl.edu
|
| Phone |
314-362-9045
|
| Organization name |
Washington University in St. Louis
|
| Department |
Pathology & Immunology
|
| Lab |
Kyunghee Choi
|
| Street address |
660S Euclid Ave.
|
| City |
St. Louis |
| State/province |
MO |
| ZIP/Postal code |
63110 |
| Country |
USA |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE45147 |
Hemangioblast skewing from embryonic stem(ES) cells by defined factors |
|
