|
| Status |
Public on Dec 31, 2011 |
| Title |
Digestive Gland-Nickel-Rep2 [MytArray 1.0] |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
digestive gland tissue, animals exposed to 0.77 mg/L Nickel for 4 days in aquaria
|
| Organism |
Mytilus galloprovincialis |
| Characteristics |
gender: female
tissue: digestive gland tissue
agent: Nickel
dose: 0.77 mg/L
time: 4 days
|
| Treatment protocol |
animals exposed in aquaria
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted from pools made of 6 different digestive gland pieces using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
15 µg of total RNA were primed with 0.5 µg oligodT(19)VN primer at 70°C for 10 min, then reversed transcribed at 42°C for 2 h in the presence of 400 U ReverseAid MuLV H minus RTase (Fermentas), and 100 µM each dATP, dTTP, dGTP, with 25 µM dCTP, 25 µM Cy3-labeled dCTP or Cy5-label dCTP
|
| |
|
| Channel 2 |
| Source name |
digestive gland tissue,vehicle treated (0.02% DMSO) for four days
|
| Organism |
Mytilus galloprovincialis |
| Characteristics |
gender: female
tissue: digestive gland tissue
agent: vehicle (0.02% DMSO)
time: 4 days
|
| Treatment protocol |
animals exposed in aquaria
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted from pools made of 6 different digestive gland pieces using Trizol following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
15 µg of total RNA were primed with 0.5 µg oligodT(19)VN primer at 70°C for 10 min, then reversed transcribed at 42°C for 2 h in the presence of 400 U ReverseAid MuLV H minus RTase (Fermentas), and 100 µM each dATP, dTTP, dGTP, with 25 µM dCTP, 25 µM Cy3-labeled dCTP or Cy5-label dCTP
|
| |
|
| |
| Hybridization protocol |
43°C, 16/20 h, hybridization buffer Northern Max (Ambion). Pre-hybridization for 1h in same conditions.
|
| Scan protocol |
The BioRad Chip Reader was used to generate TIFF files at 5 micron resolution. Then the digital images were processed using the Axon GenePix software to obtain gpr files. Gpr files were saved as gpr 3.0.
|
| Description |
n/a
|
| Data processing |
Kooperberg background subtracted, loess normalized. Data obtained from log2 of processed Ch1 signal/processed Ch2 signal. LIMMA software was used
|
| |
|
| Submission date |
Apr 06, 2010 |
| Last update date |
Dec 31, 2011 |
| Contact name |
Francesco Dondero |
| E-mail(s) |
francesco.dondero@mfn.unipmn.it
|
| Organization name |
University of Piemonte Orientale "Amedeo Avogadro"
|
| Department |
Department of Environmental and Life Sciences
|
| Lab |
Molecular Ecotoxicology
|
| Street address |
Via Michel 11
|
| City |
Alessandria |
| ZIP/Postal code |
15121 |
| Country |
Italy |
| |
|
| Platform ID |
GPL1799 |
| Series (1) |
| GSE21229 |
Mussel (Mytilus galloprovincialis) digestive gland tissue: Control vs. Nickel, Chlorpyrifos and their mixture. |
|
