|
| Status |
Public on May 05, 2012 |
| Title |
Neural tube electroporated with NEUROG2-GFP, 6h, biological rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Neural tube, overexpression of NEUROG2
|
| Organism |
Gallus gallus |
| Characteristics |
developmental stage: E1.5 (HH11)
tissue: neural tube
treatment: NEUROG2 overexpression
|
| Treatment protocol |
HH10 embryos were electroporated in ovo in the neural tube with a construct co-expressing the appropriate protein and GFP. Embryos were harvested 6h later, and for each biological replicate, neural tubes from 20 embryos were pooled prior to cell dissociation. Neural tube explants were incubated 10' in and then dissociated mechanically. GFP+ cells were collected in Tri-reagent by FACS sorting.
|
| Growth protocol |
Fertilized eggs were grown at 37°C during about 40 hours in order to reach the HH10 stage (E1.5). All of the embryos used for this study had between 13 and 15 somites developed at the time of the electroporation.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Tri-reagent extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
Biotin
|
| Label protocol |
Biotinylated cRNAs were prepared from 50 ng of total RNA using the Ambion MessageAmpTM Premier RNA Amplification Kit from Applied Biosystems according to the manufacturer's instructions.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip® Chicken Genome arrays (Affimetrix). Each microarray was then washed and stained on a GeneChip® Fluidics Station 450.
|
| Scan protocol |
Microarrays were scanned with a GeneChip® Scanner 3000 7G.
|
| Description |
SBV_Ngn1 Signal (log)
RNA extracted from FACS-sorted GFP cells.
|
| Data processing |
GeneChip Operating Software (GCOS) version 1.4 (Affymetrix, Santa Clara, CA, USA). CEL files were further processed with the Robust Multi-array Average (RMA) algorithm using the affy package in Bioconductor version release (2.4.1).
|
| |
|
| Submission date |
May 04, 2012 |
| Last update date |
May 05, 2012 |
| Contact name |
Sophie Bel Vialar |
| E-mail(s) |
sophie.vialar@univ-tlse3.fr
|
| Phone |
(33) 05 61 55 67 39
|
| Fax |
(33) 05 61 55 65 07
|
| Organization name |
CNRS
|
| Department |
UMR5567
|
| Lab |
Centre de Biologie du developement
|
| Street address |
118 rte de narbonne
|
| City |
Toulouse |
| ZIP/Postal code |
31062 |
| Country |
France |
| |
|
| Platform ID |
GPL3213 |
| Series (1) |
| GSE37782 |
Expression data from chick embryos overexpressing NEUROG2 or NEUROG2AQ in the neural tube |
|
