|
| Status |
Public on Jan 01, 2006 |
| Title |
fibroblast_cornea_control_chipB_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
fibroblast_cornea_control
|
| Organism |
Mus musculus |
| Characteristics |
Primary fibroblast, fetal mouse cornea, unstimulated control
|
| Treatment protocol |
Embryos used for fibroblast isolation were obtained from a time mated CD1 mouse. At 19-days post conception, the pregnant mouse was euthanized by CO2 asphyxiation and immediately swabbed with 70% ethanol in a sterile hood. Tendon, corneal and skin fibroblasts were isolated according to Spector et al {Spector, Goldman Robert D, et al. 1998 381 /id}. Cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 15% FCS in a 5% CO2 humidified atmosphere at 37 °C, and were subcultured when they reached approximately 80% confluence. Cells were cultured until five population doublings, at which point they were seeded onto a gelatin-coated (1% w/v) glass plate (7cm x 10cm) and allowed to adhere for approximately seven hours. STATIC CONTROL: The fibroblast-seeded glass plate was transferred to a single-well plate and overlaid with DMEM supplemented with 2% FCS and 1% penicillin/streptomycin. Fibroblasts remained under these conditions for the duration of the 14 hour experiment
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA extractions were carried out with the Absolutely RNA RT-PCR Miniprep kit (Stratagene, La Jolla, CA, USA) according to the manufacturer's instructions. Contaminating RNases were inactivated in solutions upon treatment with 0.1% (v/v) DEPC, while RNaseZap was used according to the manufacturer's instructions to eliminate RNase contamination on work surfaces and pipettes.
|
| Label |
biotin
|
| Label protocol |
Standard Affymetrix GeneChip Eukaryotic Two-Cycle Target Labeling
|
| |
|
| Hybridization protocol |
standard Affymetrix procedures
|
| Scan protocol |
standard Affymetrix procedures
|
| Description |
Primary fibroblast from the cornea of the fetal mouse, not mechanically stimulated
|
| Data processing |
Robust Multichip Average (RMA, module Affy 1.4.32 of Bioconductor)
|
| |
|
| Submission date |
Oct 20, 2005 |
| Last update date |
Oct 20, 2005 |
| Contact name |
Steve Winder |
| E-mail(s) |
s.winder@sheffield.ac.uk
|
| Phone |
+44 114 222 2332
|
| Fax |
+44 114 276 5413
|
| URL |
http://www.shef.ac.uk/bms/research/winder
|
| Organization name |
University of Sheffield
|
| Department |
Department of Biomedical Science
|
| Street address |
Western Bank
|
| City |
Sheffield |
| ZIP/Postal code |
S10 2TN |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL340 |
| Series (1) |
| GSE3486 |
Mechanically stimulated fibroblast from different fetal mouse tissues using Affy MOE430 chip set |
|
