|
| Status |
Public on Sep 14, 2013 |
| Title |
1705_383_0hr |
| Sample type |
RNA |
| |
|
| Source name |
hOB1705_shRB1383_0hr
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: Normal bone cell line 1705
phenotype: RB1 knockdown
cell type: human osteoblast (hOB)
|
| Treatment protocol |
Subconfluent hOB shEV and shRB1 cells were irradiated at 4 Gy using a 137Cesium source at a dose rate of 1.7 min/Gy and cell pelllets collected 0, 2, 4, 8, 16 and 24 hrs after irradiation
|
| Growth protocol |
Cells were maintained at 37°C and 5% CO2 in alpha MEM (modified Eagles's medium) containing 10% (v/v) FCS, 50 U.mL-1 penicillin and 50 ug.mL-1 streptomycin, 2mM L-glutamine, 100 uM L-ascorbate-2-phosphate
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from cell pellets using Trizol reagent (Invitrogen) and RNAeasy isolation kit (Qiagen). The quality of RNA was measured using Bioanalyzer (Agilent Technologies).
|
| Label |
biotin
|
| Label protocol |
Affymetrix Gene Chip® WT terminal labelling and hybridization was performed according to manufacturers instructions (Affymetrix , Santa Clara, CA).
|
| |
|
| Hybridization protocol |
Labelled RNA was hybridised to Human Gene 1.0 ST arrays. Hybridisation was performed according to the manufacturers instructions (Affymetrix , Santa Clara, CA).
|
| Scan protocol |
Arrays were washed using Affymetrix fluidics stations and scanned using the Gene Chip Scanner 3000.
|
| Description |
RB1-383 pGIPZ (OpenBiosystems)(shRB1), human osteoblast cell line (hOB)
|
| Data processing |
The data were RMA normalised using the R package Affy
|
| |
|
| Submission date |
Sep 03, 2013 |
| Last update date |
Sep 14, 2013 |
| Contact name |
Maya Kansara |
| E-mail(s) |
[email protected]
|
| Phone |
0396561618
|
| Organization name |
Peter MacCallum Cancer Centre
|
| Department |
Research
|
| Lab |
Sarcoma Genomics and Genetics
|
| Street address |
St Andrews Place
|
| City |
Melbourne |
| State/province |
Victoria |
| ZIP/Postal code |
3002 |
| Country |
Australia |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE50532 |
Gene expression analysis of RB1 knockdown in bone in response to radiation |
|
