|
| Status |
Public on Mar 25, 2014 |
| Title |
Primary calvarial osteoblast,Transfected with SC,RNA extracted 48 hours post-transfection,rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Primary calvarial osteoblast,Transfected with SC,RNA extracted 48 hours post-transfection,rep1
|
| Organism |
Mus musculus |
| Characteristics |
gender: Mixed Gender
strain: C57BL6/J
cell type: primary calvarial osteoblasts
|
| Treatment protocol |
siRNA transfection experiments cells were handled as outlined above except they were plated at 150,000 cells/well, transfected ~24h after plating using Lipofectamine 2000 Reagent (Invitrogen) according to manufacturer’s directions
|
| Growth protocol |
Primary calvarial osteoblasts were isolated from 3-9 day old neonates using sequential Collagenase P digestions. Cells were plated into 6 well plates at 300,000 cells/2ml sterile plating media (DMEM, 10% heat-inactivated FBS, 100 U/ml penicillin, 100 μg/ml streptomycin) per well.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the Qiagen Rneasy Mini Kit.
|
| Label |
Biotin
|
| Label protocol |
Biotin-labeled cRNA was synthesized by the total prep RNA amplification kit from Ambion (Austin, TX).
|
| |
|
| Hybridization protocol |
cRNA was quantified and normalized to 77 ng/μl, and then 850 ng was hybridized to Beadchips.
|
| Scan protocol |
The hybridized Beadchips were scanned by Illumina BeadScan confocal scanner. cRNA synthesis, hybridization and scanning was performed by the Genome Sciences Laboratory in the Center for Public Health Genomics at the University of Virginia.
|
| Data processing |
Illumina GenomeStudio generated raw intensity values. The expression values were transformed using the Variance Stabilizing Transformation (VST), and quantile normalized using the LumiR R package.
|
| |
|
| Submission date |
Oct 25, 2013 |
| Last update date |
Mar 25, 2014 |
| Contact name |
Charles R Farber |
| E-mail(s) |
crf2s@virginia.edu
|
| Organization name |
University of Virginia
|
| Department |
Center for Public Health Genomics
|
| Street address |
P.O. Box 800717
|
| City |
Charlottesville |
| State/province |
VA |
| ZIP/Postal code |
22908 |
| Country |
USA |
| |
|
| Platform ID |
GPL6885 |
| Series (1) |
| GSE51693 |
Osteoblast Gene Expression in Response to Bicc1 Knockdown |
|
