|
| Status |
Public on Jan 18, 2014 |
| Title |
E14.5 B |
| Sample type |
RNA |
| |
|
| Source name |
Scx-positive tendon cells from Scx-GFP mice
|
| Organism |
Mus musculus |
| Characteristics |
cell: limb tendon cells
developmental stage: E14.5
|
| Treatment protocol |
Forelimbs from E11.5, E12.5 and E14.5 Scx-GFP embryos were collected and dissociated with trypsin to obtain cell suspensions. Cell suspensions were subjected to FACS at room temperature using a MoFlo® XDP Flow Cytometer (Dako, with state laser 488 nm) with the Dako-Moflo Summit software or using a VantageTM Se option DiVa flow cytometer (laser 488 nm). The GFP-positive fractions were collected in PBS-EDTA 2 mM supplemented with 20% of foetal calf serum. RNA quantity was monitored on Agilent RNA Pico LabCHips (Agilent). Fragmented biotin-labelled cRNA samples were hybridized on Affymetrix Gene Chip Mouse Genome 430 2.0 arrays
|
| Growth protocol |
Scx-GFP mouse embryos were collected at E11.5, E12.5 or E14.5 after natural overnight mattings. For staging, fertilization was considered to take place at 0 a.m..
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA were extract with with the Qiagen RNeasy minikit (Qiagen), then 100 ng of total RNA were amplified using GeneChip 3' IVT Express Kit from Affymetrix.
|
| Label |
biotin
|
| Label protocol |
As per manufacturer (Affymetrix)
|
| |
|
| Hybridization protocol |
As per manufacturer (Affymetrix)
|
| Scan protocol |
The Arrays were scanned using the Gene Chip Scanner 3000. Then CEL files were generated using the Affymetrix GeneChip Command Console software (AGCC) 3.0
|
| Data processing |
CHP files were processed using MAS 5.0 analysis algorithm with Affymetrix Expression Console software 1.1
|
| |
|
| Submission date |
Jan 17, 2014 |
| Last update date |
Jan 18, 2014 |
| Contact name |
Delphine Duprez |
| E-mail(s) |
[email protected]
|
| Phone |
+33 1 44 27 27 53
|
| Organization name |
CNRS/UPMC
|
| Department |
UMR 7622
|
| Lab |
Developmental Biology
|
| Street address |
9, Quai Saint Bernard
|
| City |
Paris |
| ZIP/Postal code |
75005 |
| Country |
France |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE54207 |
Expression data from mouse limb tendon cells during development. |
|
