|
| Status |
Public on Jul 08, 2010 |
| Title |
72 hr |
| Sample type |
RNA |
| |
|
| Source name |
72 hr after mechanical stress
|
| Organism |
Rattus norvegicus |
| Characteristics |
strain: Sprague-Dawley
age: E15
tissue: cultured spinal cord cells
|
| Treatment protocol |
Dissected tissues were rinsed with cold Ca2+ -and Mg2+-free Hanks balanced salt solution (HBSS) supplemented with 4 g/L glucose, and incubated at 37°C for 20 minutes with 0.03% (w/v) trypsin solution in HBSS under mild shaking.
|
| Growth protocol |
The spinal cords of Sprague-Dawley rat embryos were dissected out at post-coital day 15 and stripped of the dorsal root ganglia and meninges.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The cultured cells on each well at 0, 2, 6, 12, 24, 48 and 72 hours were disrupted in a lysis buffer containing β-mercaptoethanol and the total RNA from five animals was pooled at each time point, and further purified using RNeasy® Mini Kit (Qiagen, Valencia, CA) and treated with DNase I (TaKaRa Bio, Ohtsu, Japan)
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were synthesized using the IVT Labeling Kit (Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 20 µg of cRNA were hybridized for 16 hours at 45°C on the GeneChip® Rat Genome 230 2.0 Array.
|
| Scan protocol |
GeneChips were washed and stained in the Affymetrix Fluidics Station 450, and scanned using GeneChip Scanner 3000 7G.
|
| Description |
Gene expression 72 hr after stress in E15 Sprague-Dawley rats spinal cord
|
| Data processing |
Microarray data was initially processed using GeneChip Operating Software (GCOS, Affymetrix). Signal intensity was calculated by Microarray Suite version 5.0 (MAS5.0) with Affymetrix default setting and global scaling as the normalization method.
|
| |
|
| Submission date |
Jul 07, 2010 |
| Last update date |
Jul 07, 2010 |
| Contact name |
Kenzo Uchida |
| E-mail(s) |
kuchida@u-fukui.ac.jp
|
| Phone |
81-776-61-8383
|
| Fax |
81-776-61-8125
|
| Organization name |
Fukui University Faculty of Medical Sciences
|
| Department |
Department of Orthopaedics and Rehabilitation Medicine
|
| Street address |
Shimoaizuki 23
|
| City |
Matsuoka |
| State/province |
Fukui |
| ZIP/Postal code |
910-1193 |
| Country |
Japan |
| |
|
| Platform ID |
GPL1355 |
| Series (1) |
| GSE22791 |
Microarray analysis of expression of cell death-associated genes in rat spinal cord cells exposed to cyclic tensile stresses in vitro |
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