|
| Status |
Public on Nov 17, 2010 |
| Title |
7 Days-3-2 |
| Sample type |
RNA |
| |
|
| Source name |
embryoid bodies
|
| Organism |
Mus musculus |
| Characteristics |
developmental stage: 7 days
cell type: embryoid bodies
|
| Treatment protocol |
Mouse stem cells were differentiated spontaneously in cell culture to form Ebs
|
| Growth protocol |
MES3 mESC line was derived by whole blastocyst culture from the inner cell mass of a C57BLxCBA mouse embryo. Culture cells were maintained on Mitomicin C inactivated mouse embryonic fibroblasts (MEF) in knock-out DMEM (Gibco, Invitrogen) supplemented with 10% FBS (Gibco, Invitrogen), 2mM non-essential amino acids, 2 mM L-glutamine, 50 mg/mL Penn-Strep, 10 nM b-mercaptoethanol and 1000 units/mL LIF (ESGRO, Chemicon). EBs were obtained by excising clusters of undifferentiated stem cells from growing colonies and transferring them to low-adhesion culture plates, where they were maintained in the same medium as the mESCs but without b-mercaptoethanol and LIF.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from ESC cultures by the Trizol® method (Invitrogen). All samples were treated with an RNase-free DNase set (Qiagen, Valencia, CA)
|
| Label |
Cy5
|
| Label protocol |
linear amplification method using Ambion’s MessageAmp™ II‐Biotin Enhanced Single Round aRNA Amplification Kit
|
| |
|
| Hybridization protocol |
Amersham Biosciences CodeLink protocol (Ramakrishnan et al, 2002)
|
| Scan protocol |
Axon GenePix 4000B scanner (Axon Instruments, Foster City, CA) with 10 M resolution. CodeLink™ Expression Analysis software v 5.0
|
| Description |
T00340101
|
| Data processing |
Normalization: 1) Background correction: saddle-point approximation (offset = 50); 2) quantile normalization method
|
| |
|
| Submission date |
Nov 16, 2010 |
| Last update date |
Nov 16, 2010 |
| Contact name |
Angel Concha |
| E-mail(s) |
angel.concha.sspa@juntadeandalucia.es
|
| Organization name |
Servicio Andaluz de Salud
|
| Street address |
2 Fuerzas Armadas
|
| City |
Granada |
| ZIP/Postal code |
18014 |
| Country |
Spain |
| |
|
| Platform ID |
GPL2897 |
| Series (1) |
| GSE25403 |
Genome-wide gene expression analysis in mouse embryonic stem cells |
|
