|
| Status |
Public on Aug 04, 2013 |
| Title |
Parental clone +Doxycycline, day2, amp, rep2 |
| Sample type |
RNA |
| |
|
| Source name |
BVSCR26rtTA ESCs +Doxycycline, day2
|
| Organism |
Mus musculus |
| Characteristics |
gender: Male
genetic background: C57BL/6 + 129
transgene: Blimp1-mVenus, stella-ECFP, Rosa26-rtTA
cell type: Parental ESCs clone
|
| Treatment protocol |
After 36 hrs of differentiation, cells were harvested and cultured in a Lipidure-Coat 96-well plate (NOF) to be aggregated (started with 2,000 cells/well) in GK15 with 1.5 μg/ml of Dox (Clonetech) to induce TF-PGCLCs. Ck-PGCLCs were induced by BMP4 (500 ng/ml), BMP8A (500 ng/ml), SCF (100 ng/ml), LIF (1000 U/ml) and EGF (50 ng/ml) as previously described (Hayashi et al., Cell, 2011)
|
| Growth protocol |
Transfected ESCs were maintained under 2i+LIF condition and adapted to a feeder-free condition prior to induction. EpiLC differentiation was performed as reported previously (Hayashi et al., Cell, 2011).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
total RNA was extracted using Qiagen RNeasy according to manufacturer's instruction
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared from total RNA according to the standard Affymetrix protocol (Affymetrix GeneChip Expression Analysis Technical Manual, 2004, Affymetrix: Eukaryotic One-Cycle Target Labeling Assay)
|
| |
|
| Hybridization protocol |
Hybridization was performed according to the standard Affymetrix protocol (Affymetrix GeneChip Expression Analysis Technical Manual, 2004, Affymetrix: Eukaryotic Target Hybridization)
|
| Scan protocol |
The microarray image data were processed with the GeneChip Scanner 3000 (Affymetrix)
|
| Description |
Male, C57BL/6;129, Blimp1-mVenus, stella-ECFP, Rosa26-rtTA
|
| Data processing |
CEL data were generated by GeneChip Operating Software and then subjected to the dCHIP software. Data were normalized together with the default settings.
|
| |
|
| Submission date |
Jan 25, 2013 |
| Last update date |
Aug 04, 2013 |
| Contact name |
Fumio NAKAKI |
| E-mail(s) |
[email protected]
|
| Phone |
+81-75-753-4343
|
| Fax |
+81-75-751-7286
|
| Organization name |
Kyoto University
|
| Department |
Graduate School of Medicine
|
| Lab |
Anatomy and Cell Biology
|
| Street address |
Yoshidakonoe-cho, Sakyo-ku
|
| City |
Kyoto |
| ZIP/Postal code |
606-8501 |
| Country |
Japan |
| |
|
| Platform ID |
GPL1261 |
| Series (2) |
| GSE43775 |
Induction of the mouse germ cell fate by transcription factors in vitro [exp1] |
| GSE46855 |
Induction of the mouse germ cell fate by transcription factors in vitro |
|
