|
| Status |
Public on Sep 02, 2008 |
| Title |
Post-irradiated at 2.0Gy 2hrs Rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Foreskin fibroblasts (AG1522)
|
| Organism |
Homo sapiens |
| Characteristics |
Low-LET irradiation of 2.0 Gy
Expression after 2hrs
|
| Treatment protocol |
Confluent cell cultures were irradiated with 250 kV X-rays at two dose levels, either 0.1 or 2 Gy, at NASA’s Johnson Space Center (JSC, Houston, TX, USA). The exposure at 2 Gy took 3.2 minutes; that at 0.1 Gy took 42 seconds.
|
| Growth protocol |
Cells and culture conditions were previously described by Wu et al. (2002) Journal of radiation research 43 Suppl:S181-5.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted by Trizol reagent.
|
| Label |
[32P]dCTP
|
| Label protocol |
Three µg of total RNA was reverse-transcribed using oligo-dT primers (Invitrogen) and M-MLV reverse transcribed (Invitrogen) in the presence of 100 mCi of [32P]dCTP (Amersham Bioscience, Piscataway, NJ) according to Maes et al. 2007 (Neurobiol Aging. 28:1795-809).
|
| |
|
| Hybridization protocol |
Nadon NL, Mohr D, Becker KG. National Institute on Aging microarray facility--resources for gerontology research. Journals of Gerontology Series A-Biological Sciences & Medical Sciences 2005;60(4):413-5 (PMID: 15933377)
|
| Scan protocol |
Hybridization intensities were measured using a Storm PhosphorImager (Amersham BioSciences, USA).
|
| Description |
A broad coverage of the human genome (approximately 6424 unique genes) was analyzed using the NIA Human MGC cDNA microarray, which consist of full-length cDNA clones generated by the National Cancer Institute’s Mammalian Gene Collection (http://mgc.nci.nih.gov/).
|
| Data processing |
Data acquisition and global background correction were obtained using the Array-Pro Analyzer 4.5 software (Media Cybernetics, MD, USA). There has been no data processing or normalization of the background-corrected raw data prior to their analysis.
Net intensity was derived from whole cell area measurement and corrected with mean intensity from background cells, calculated using a biquadratic polynomial.
|
| |
|
| Submission date |
Sep 02, 2008 |
| Last update date |
Sep 02, 2008 |
| Contact name |
Olivier Charles Maes |
| E-mail(s) |
oliviercharlesmaes@yahoo.ca
|
| Phone |
514-340-8222
|
| Fax |
514-340-8295
|
| Organization name |
Lady Davis Institute for Medical Research
|
| Department |
Bloomfield Centre for Research in Aging
|
| Lab |
428
|
| Street address |
3999 Cote Ste Catherine
|
| City |
Montreal |
| State/province |
Quebec |
| ZIP/Postal code |
H3T 1E2 |
| Country |
Canada |
| |
|
| Platform ID |
GPL1211 |
| Series (1) |
| GSE12647 |
Gene expression profiling in human fibroblast after low-LET irradiation |
|
