|
| Status |
Public on Dec 01, 2010 |
| Title |
Mouse ES 1 [U74Cv2] |
| Sample type |
RNA |
| |
|
| Source name |
Mouse ES cells (129 derived)
|
| Organism |
Mus musculus |
| Characteristics |
culture cells: Mouse ES cells
sorted for: E-cadherin+
|
| Growth protocol |
Mouse Sox1-GFP ES cells (129 derived) were maintained as described (http://www.ncbi.nlm.nih.gov/pubmed/15696161). The differentiation medium was GMEM supplemented with 10% Knockout Serum Replacement (KSR; Invitrogen), 2 mM glutamine, 1 mM pyruvate, 0.1 mM nonessential amino acids, and 0.1 mM 2-ME with or without 10% fetal calf serum (FCS; JRH). For the SFEBculture, ES cells were dissociated to single cells in 0.25% trypsin-EDTA (Invitrogen) and reaggregated in the differentiation medium (5*10^5 cells/dish) using bacterial grade dish.
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
RNA was isolated from 1*10^6 cells of FACS sorted population with RNeasy Mini Kit (Qiagen) according to the manufacturer’s instructions. PolyA RNA was annealed by T7-dT primer and cDNA was synthesised by Superscript II (Invitrogen). cDNA was amplifed by T4 DNA polymerase and purified with Ph/Ch method.
|
| Label |
biotin
|
| Label protocol |
Biotin labeled cRNA was synthesised by T7 RNA polymerase with Rnase inhibitor for 4hr at 37℃. RNA was purified by RNeasy Mini Kit.
|
| |
|
| Hybridization protocol |
Samples were hybridized with Affymetrix Murine Genome U74C Version 2 Array and scanned at the RIKEN CDB Microarray Facility.
|
| Scan protocol |
Array scanning was performed according to the manufacturer's instruction (Affymetrix)
|
| Data processing |
The expression data were analyzed using an in-house-developed program (eXintegrator, http://www.cdb.riken.jp/scb/documentation/index.html). eXintegrator is a program that converts processed/normalized CHP data into the visual graphic. Therefore, final processed/normalized data is CHP in our study. Algorithm: ExpressionStat 5.0.
|
| |
|
| Submission date |
Nov 30, 2010 |
| Last update date |
Dec 08, 2010 |
| Contact name |
Daisuke Kamiya |
| E-mail(s) |
kamiya@cdb.riken.jp
|
| Organization name |
RIKEN CDB
|
| Street address |
2-2-3 Minatojima-Minamimachi, Chuo
|
| City |
Kobe |
| ZIP/Postal code |
6500047 |
| Country |
Japan |
| |
|
| Platform ID |
GPL83 |
| Series (1) |
| GSE25593 |
Expression Profiling: during in vitro neural differentiation from mES cells. |
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