|
| Status |
Public on Nov 30, 2017 |
| Title |
KO3 |
| Sample type |
RNA |
| |
|
| Source name |
PGCs from KO E13.5 embryos
|
| Organism |
Mus musculus |
| Characteristics |
strain/genotype: og2.fvb.fancg -/-
developmental stage: E13.5
gender: male
cell type: Primordial Germ Cells
|
| Treatment protocol |
Gonads were removed from E13.5 embryos and lysed 5 minutes at 37°C to obtain a single cells suspension, using a dissociation buffer containing PBS1X without Ca2+ and Mg2+, collagenase (final concentration 1 mg/ml), and DNase I (final concentration 0.02 mg/ml).
|
| Growth protocol |
og2.fvb.fancg -/- and og2.fvb.fancg +/+ mice were used to sort PGCs
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cell suspension was analysed by flow cytometry and PGCs sorted according to GFP fluorescence. Sorted PGC were retrieved in the lysis buffer from the the RNeasy micro kit from Qiagen and RNA repared according to the manufacturer's instructions
|
| Label |
biotin
|
| Label protocol |
Biotinylated cDNA were prepared according to the standard Nugen Ovation PicoV2 protocol from 500 pg total RNA.
|
| |
|
| Hybridization protocol |
Following fragmentation, 3,6 ug of cDNA were hybridized for 17 hr at 45C on GeneChip MouseGene2.0ST Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using Affymetrix GC3000 Scanner.
|
| Description |
Gene expression data from Fancg-/- E13.5 male Primordial Germ Cells
|
| 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 |
Nov 29, 2017 |
| Last update date |
Jan 23, 2018 |
| Contact name |
Pierre Fouchet |
| E-mail(s) |
pierre.fouchet@cea.fr
|
| Organization name |
CEA
|
| Street address |
10, route du panorama
|
| City |
Fontenay aux roses |
| ZIP/Postal code |
92260 |
| Country |
France |
| |
|
| Platform ID |
GPL18802 |
| Series (1) |
| GSE107504 |
Transcriptomic comparison of E13.5 Primordial Germ Cells (PGCs) from Fancg-/- and WT mice |
|
