|
| Status |
Public on Dec 02, 2017 |
| Title |
FOG-2 Wt-18-2 |
| Sample type |
RNA |
| |
|
| Source name |
proliferating Wt MEF experiment-2
|
| Organism |
Mus musculus |
| Characteristics |
genotype: wildtype
cell type: embryonic fibroblast
developmental stage: E14.5
|
| Treatment protocol |
Growing Wt and FOG-2 Rb- MEF were trypsinized and one million of cell pellet were washed with PBS before RNA extraction
|
| Growth protocol |
Wt and FOG-2 Rb- MEF were grown in DMEM/F12 containing 20% Donor Bovine Serum, 2mM L-Glutamine, 1x Penicillin/Streptomycin and 0,1mM beta-mecaptoethanol
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA coupled to RNAeasy kit (Ambion) purification was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
270 ng of total RNA is reverse transcribed following the Genechip Whole transcript (WTplus) Sense Target labelling assay kit (Affymetrix). Briefly, the resulting double strand cDNA is used for in vitro transcription with T7 RNA pol. This cRNA is reverse transcribed in single stranded sens DNA which is fragmented and biotin labelled
|
| |
|
| Hybridization protocol |
biotynilated single stranded sense cDNA is then hybridized to GeneChip® Human Gene2.0 (Affymetrix) at 45°C for 17 hours
|
| Scan protocol |
After overnight hybridization, chips are washed on the fluidic station FS450 following specific protocols (Affymetrix) and scanned using the GCS3000 7G
|
| Data processing |
Data were normalized using RMA and statistical analysis was realized on Partek Genomics Suite. CDF file V20 was used for annotation.
|
| |
|
| Submission date |
Dec 05, 2016 |
| Last update date |
Dec 02, 2017 |
| Contact name |
Franck Letourneur |
| E-mail(s) |
franck.letourneur@inserm.fr
|
| Organization name |
INSERM
|
| Department |
U1016
|
| Lab |
genomic
|
| Street address |
22 rue mechain
|
| City |
Paris |
| ZIP/Postal code |
75014 |
| Country |
France |
| |
|
| Platform ID |
GPL22598 |
| Series (1) |
| GSE90859 |
Study of knock-in mice expressing the co-Factor FOG-2 that can not bind pRb |
|
