|
| Status |
Public on Sep 17, 2020 |
| Title |
SR1FP_III_RC4_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
foot pad cells after first transplantation into skin, bulk
|
| Organism |
Rattus norvegicus |
| Characteristics |
genotype: constitutively express the Enhanced Green Fluorescent protein (EGPF) (green rat CZ-004” SDTgN (act-EGFP)obsCZ-004)
background strain: Sprague-Dawley
tissue: foot pad
|
| Treatment protocol |
Tissues and hair follicle cells were cultivated on a feeder layer of irradiated 3T3-J2 cells in a 3:1 DMEM F12 medium supplemented as described (Bonfanti et al., 2010). EGFP-positive cells were FACS-sorted for analysis.
|
| Growth protocol |
Various tissues were dissected from PN200 EFGP rat. Dissected tissues were harvested and kept in cold Dulbecco Modified Eagle's Medium (DMEM) supplemented with 10% Bovine Serum during dissection. Tissues were mechanically minced before being dissociated in 0.05% trypsin solution containing 0.1% EDTA at 37°C with a gentle stirring. Skin grafts containing GFP-positive hair follicles were carefully dissected from the back of recipient nude mice and dissociated in 30 ml of 0.05% trypsin containing 0.1% EDTA at 37°C for 60 to 120 min.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was prepared using the RNeasy Mini Kit (QIAGEN Inc.)
|
| Label |
biotin
|
| Label protocol |
100 nanogram of total RNA were amplified and labeled using Affymetrix whole transcript sense target labeling protocol (kits #900673, #900671)
|
| |
|
| Hybridization protocol |
Affymetrix Rat Gene 1.0 ST arrays were hybridized with 5 microgramme of labeled, amplified cDNA , washed, stained and scanned according to the protocol described in Affymetrix GeneChip® Expression Analysis Manual (Fluidics protocol FS450_0007)
|
| Scan protocol |
Scanning was done on an Affymetrix GeneChip Scanner 7G
|
| Description |
PB_42
|
| Data processing |
Normalized expression signals were calculated from Affymetrix CEL files using RMA (Irizarry et al., 2003) in Affymetrix Expression Console software.
|
| |
|
| Submission date |
Jul 06, 2018 |
| Last update date |
Sep 17, 2020 |
| Contact name |
Sandra Calderon |
| Organization name |
University of Lausanne
|
| Department |
Center for Integrative Genomics
|
| Lab |
Lausanne Genomics Technologies Facility
|
| Street address |
Génopode Building
|
| City |
Lausanne |
| ZIP/Postal code |
1015 |
| Country |
Switzerland |
| |
|
| Platform ID |
GPL6247 |
| Series (1) |
| GSE116719 |
Comparison of epithelial cells from various epithelia and hair follicle stem cells [microarray] |
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