|
| Status |
Public on Feb 11, 2016 |
| Title |
CD45-Ter119-ItgavlowTie2+rep3 |
| Sample type |
RNA |
| |
|
| Source name |
wild-type, anterior posterior limb
|
| Organism |
Mus musculus |
| Characteristics |
age: postnatal day 3
tissue: fore and hind limbs
genotype: wild type
|
| Treatment protocol |
none
|
| Growth protocol |
Skeletal tissues were dissected from P3 GFP-labeled mice and dissociated by mechanical and enzymatic dissociation. Specifically, the tissue was placed in collagenase digestion buffer supplemented with DNase and incubated at 37oC for 40 minutes under constant agitation. After collagenase digestion and neutralization, undigested materials were gently triturated by repeated pipetting. Total dissociated cells were filtered through 40 m nylon mesh, pelleted at 200g at 4oC, resuspended in staining media (2% fetal calf serum in PBS), blocked with rat IgG and stained with fluorochrome-conjugated antibodies against CD45, Tie2, AlphaV integrin, CD105, Thy1.1, Thy 1.2, 6C3 and CD200 for fractionation by fluorescence activated-cell sorting.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was isolated with RNeasy Micro Kit (Qiagen, Germantown, MD) as per manufacturer’s instructions. RNA was twice amplified with a RiboAmp RNA amplification kit (Arcturus Engineering, Mountain View, CA).
|
| Label |
biotin
|
| Label protocol |
Amplified cRNA was streptavidin-labeled, fragmented, and hybridized to Affymetrix 430-2.0 arrays as recommended by the manufacturer (Affymetrix, Santa Clara). Arrays were scanned with a Gene Chip Scanner 3000 (Affymetrix) running GCOS 1.1.1. software.
|
| |
|
| Hybridization protocol |
standard Affymetrix protocol
|
| Scan protocol |
Arrays were scanned with a Gene Chip Scanner 3000 (Affymetrix) running GCOS 1.1.1. software.
|
| Description |
fore and hind limbs pooled litter
|
| Data processing |
Normalized Intenstity values were computed based on Gene Expression Commons Algorithum [ https://gexc.stanford.edu/]
|
| |
|
| Submission date |
Jan 21, 2016 |
| Last update date |
Feb 11, 2016 |
| Contact name |
Rahul Sinha |
| E-mail(s) |
[email protected]
|
| Organization name |
Stanford University School of Medicine
|
| Department |
ISCBRM
|
| Lab |
Weissman
|
| Street address |
265 Campus Drive
|
| City |
Stanford |
| State/province |
CA |
| ZIP/Postal code |
94305 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
|
