|
| Status |
Public on Feb 20, 2012 |
| Title |
ER71 overexpressed Flk-1 mesoderm |
| Sample type |
RNA |
| |
|
| Source name |
No doxycycline treated iER71, ES cell
|
| Organism |
Mus musculus |
| Characteristics |
age: Day 3
tissue: embryonic stem cell
genotype: wild-type
doxycycline treated: no
|
| 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 into the GeneChipHybridization Oven 640 for 18 hr at 45C on Exon 1.0 ST Array. GeneChip were washed and stained in the Affymetrix Fluidics Station 450
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip 7G 3000
|
| Description |
Flk-1+ cells sorted from day3 differentiated ES cells
|
| Data processing |
Data were processed using Affymetrix Expression Console software. Gene level expression were summarized from the intensities and then normalized using the RMA algorithm.
probe group file: MoEx-1_0-st-v1.r2.pgf
meta-probeset file: MoEx-1_0-st-v1.r2.dt1.mm8.mps
|
| |
|
| Submission date |
Feb 13, 2012 |
| Last update date |
Nov 15, 2012 |
| 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 |
GPL6096 |
| Series (1) |
| GSE35793 |
Expression data from day 3 Flk-1 mesoderm between ER71 overexpressed and ER71 knockout mouse embryonic stem cell |
|
