|
| Status |
Public on Sep 04, 2012 |
| Title |
Kidney B Lgr5 pos |
| Sample type |
RNA |
| |
|
| Source name |
FACS-sorted cells from cultured embryonic kidneys from Lgr5-EGFP-Ires-CreERT2 mice, Lgr5 positive
|
| Organism |
Mus musculus |
| Characteristics |
strain: B6.129P2-Lgr5tm1(cre/ERT2)Cle/J
tissue: embryonic kidney
lgr5 expression level: positive
|
| Treatment protocol |
Embryonic kidneys were identified by fluorescence microscopy and enzymatically digested to a single cell solution using TrypLE (Sigma Aldrich) and DNase.
|
| Growth protocol |
Murine embryonic kidneys were isolated and cultured as previously described (Giuliani, Journal of Urology 2008 (PMID 18006007)).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the RNeasy Mini Kit (Qiagen) following the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
WT labeling kit (www.affymetrix.com).
|
| |
|
| Hybridization protocol |
Biotin terminal labeled sense strand DNA was generated from 100 ng total RNA using the Ambion WT Expression Kit (Applied Biosytems, 4425209 Rev. C), followed by end labeling using the Affymetrix GeneChip WT Terminal Labeling Kit conforming to the manufacturer's instructions (P/N 702808 Rev. 4). Final yields were measured using a Nanodrop 1000 (v.3.8.0, Thermo Scientific). The labeled samples were hybridized on Affymetrix Mouse Gene 1.1 ST arrays using a GeneChip Hybridisation Oven 640. Chips were labeled and washed using the GeneChip Hybridisation, Wash, and Stain Kit (Affymetrix) in a GeneChip Fluidics Station 450 (P/N 08-0092).
|
| Scan protocol |
Scanning was performed using a GeneChip Scanner 3000 7G conforming to the manufacturer's instructions (P/N 702731).
|
| Description |
ext392
Lgr5ki_GFP-positive 2.
|
| Data processing |
AffyPowerTools-rma-sketch.
|
| |
|
| Submission date |
Aug 03, 2012 |
| Last update date |
Sep 04, 2012 |
| Contact name |
Richard Volckmann |
| E-mail(s) |
r.volckmann@amsterdamumc.nl
|
| Organization name |
Academic Medical Centre, University of Amsterdam
|
| Department |
Department of Oncogenomics
|
| Street address |
P.O. Box 22700
|
| City |
Amsterdam |
| ZIP/Postal code |
1100DE |
| Country |
Netherlands |
| |
|
| Platform ID |
GPL11533 |
| Series (1) |
| GSE39881 |
Lgr5+ve stem cells in nephrogenesis |
|
