|
| Status |
Public on Dec 22, 2011 |
| Title |
N2a cells transfected by FLAG 5 |
| Sample type |
RNA |
| |
|
| Source name |
Neuroblastoma cells
|
| Organism |
Mus musculus |
| Characteristics |
tissue: Brain
cell line: N2a neuroblastoma cells
genotype/variation: control
|
| Treatment protocol |
Cells were transfected with an expression vector expressing Arx using Lipofectamine 2000 according to the guideline of the manufacturer (Invitrogen)
|
| Growth protocol |
N2a cells were grown in Dulbecco's Modified Eagle Medium (DMEM) high glucose (PAA) which was supplemented with 10% fetal bovine serum gold (PAA), 2 mM L-glutamine (Lonza), 0,1 mM non essential amino-acids (Gibco) and 2 mM penicillin/streptomycin (PAA). Cells were grown at 37°C with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Two days after transfection, RNA from N2a cells was isolated using the Total RNA Isolation Nucleospin RNA II kit following the manufacturer's instructions (Macherey-Nagel).
|
| Label |
Cy5
|
| Label protocol |
RNA was labelled using the Low Input Quick Amp Labelling kit (Agilent Technologies). The fluorescent labelled cRNA probes were purified using the RNeasy Mini kit (Qiagen).
|
| |
|
| Hybridization protocol |
825 ng of cyanine-labeled, linearly amplified cRNA were hybridized at 65°C at 10 rpm for 17 hours, with 10X Blocking Agent, 25X Fragmentation Buffer, Nuclease free water and 2X GEx Hybridization Buffer HI-RPM (Agilent Technologies). Samples were hybridized on Mouse Gene Expression 4x44K V2 microarray (Agilent Technologies).
|
| Scan protocol |
Hybridization images were obtained using an Agilent DNA microarray scanner (G2505B). Profile : Agilent HD_GX_2color / Scan Region : Agilent HD (61 x 21,6 mm) / Resolution : 5 µm / TIFF : 20 bit / RPMT : 100% / GPMT : 100%. Intensity data were extracted using Feature Extraction software (FE v9,5,1) (Agilent Technologies).
|
| Description |
FLAG-5_10142_1
|
| Data processing |
The Lowess method was used to normalize data expression (using in-house R script).
|
| |
|
| Submission date |
Jun 24, 2011 |
| Last update date |
Dec 22, 2011 |
| Contact name |
Gaelle Friocourt |
| E-mail(s) |
gaellefriocourt@yahoo.fr
|
| Phone |
+33 298443895
|
| Fax |
+33 298467910
|
| Organization name |
Inserm U613
|
| Department |
Molecular Genetics
|
| Street address |
46 rue Félix Le Dantec, CS51819
|
| City |
Brest |
| ZIP/Postal code |
29218 |
| Country |
France |
| |
|
| Platform ID |
GPL10333 |
| Series (2) |
| GSE30190 |
Comparison of gene expression between Arx-transfected N2a cells and cells transfected by the corresponding empty vector |
| GSE30191 |
ARX transcription factor implicated in mental retardation and epilepsy |
|
