|
| Status |
Public on Dec 15, 2008 |
| Title |
U2OS Dex 69th hour |
| Sample type |
RNA |
| |
|
| Source name |
U2OS osteosarcoma
|
| Organism |
Homo sapiens |
| Characteristics |
synchronized with dexamethasone
|
| Treatment protocol |
Cells were synchronized with Dexamethasone 24 hours before samples were collected.
|
| Growth protocol |
U2OS cells (ATCC) were grown to confluence in DMEM supplimented with 10% FCS and P/S/G on 35mm dishes.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were washed briefly with 1X PBS. 1 mL Trizol (Invitrogen) was used to homogenize cells, and samples were frozen at -80 until total RNA was purified.
|
| Label |
biotin
|
| Label protocol |
Second-strand cDNA synthesis was followed by in vitro transcription for linear amplification of each transcript and incorporation of biotinylated CTP and UTP.
|
| |
|
| Hybridization protocol |
The cRNA products were fragmented to 200 nucleotides or less, heated at 99 degrees C for 5 min and hybridized for 16 h at 45 degrees C. The microarrays were then washed at low (6X SSPE) and high (100mM MES, 0.1M NaCl) stringency and stained with streptavidin-phycoerythrin. Fluorescence was amplified by adding biotinylated anti-streptavidin and an additional aliquot of streptavidin-phycoerythrin stain.
|
| Scan protocol |
A confocal scanner was used to collect fluorescence signal at 3um resolution after excitation at 570 nm. The average signal from two sequential scans was calculated for each microarray feature.
|
| Description |
n/a
|
| Data processing |
signal calculated by RMA in Affymetrix Expression Console
|
| |
|
| Submission date |
Dec 15, 2008 |
| Last update date |
Dec 15, 2008 |
| Contact name |
Michael Hughes |
| E-mail(s) |
michael.evan.hughes@gmail.com
|
| Organization name |
UPenn
|
| Street address |
421 Curie Bvd, Brb 835
|
| City |
Philadelphia |
| State/province |
PA |
| ZIP/Postal code |
19104 |
| Country |
USA |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE13949 |
High temporal resolution profiling of U2OS fibroblasts |
|
