|
| Status |
Public on Oct 02, 2009 |
| Title |
IR-Vt/Mt-3 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
IR-Vt-3
|
| Organism |
Solea senegalensis |
| Characteristics |
tissue: ovary pieces
developmental stage: vitellogenic
|
| Treatment protocol |
To collect females with maturing and mature ovaries, fish were treated with intramuscular injection of 5 μg/kg GnRHa and killed 24-48 h later.
|
| Growth protocol |
Females were sacrificed at different times during the annual reproductive cycle and pieces of the ovary were deep-frozen
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from vitellogenic and mature ovaries using the RNeasy extraction kit (Qiagen) and treated with DNAse following the manufacturer’s instructions
|
| Label |
Cy5
|
| Label protocol |
Total RNA (0.5 μg) from each sample was amplified and labelled with fluorescent cyanine dyes, Cy3 or Cy5, using the Eberwein mRNA amplification procedure employing the MessageAmp™ aRNA amplification kit from Ambion (Applied Biosystems) following the manufacturer's instructions with minor modifications
|
| |
|
| Channel 2 |
| Source name |
IR-Mt-3
|
| Organism |
Solea senegalensis |
| Characteristics |
tissue: ovary pieces
developmental stage: mature
|
| Treatment protocol |
To collect females with maturing and mature ovaries, fish were treated with intramuscular injection of 5 μg/kg GnRHa and killed 24-48 h later.
|
| Growth protocol |
Females were sacrificed at different times during the annual reproductive cycle and pieces of the ovary were deep-frozen
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from vitellogenic and mature ovaries using the RNeasy extraction kit (Qiagen) and treated with DNAse following the manufacturer’s instructions
|
| Label |
Cy3
|
| Label protocol |
Total RNA (0.5 μg) from each sample was amplified and labelled with fluorescent cyanine dyes, Cy3 or Cy5, using the Eberwein mRNA amplification procedure employing the MessageAmp™ aRNA amplification kit from Ambion (Applied Biosystems) following the manufacturer's instructions with minor modifications
|
| |
|
| |
| Hybridization protocol |
Hybridizations were carried out for 17 h at 60°C using Agilent's gaskets G2534-60002, G2534A hybridization chambers, and DNA Hybridization Oven G2545A, according to the manufacturer's instructions
|
| Scan protocol |
Microarray raw data were obtained using Agilent's DNA Microarray Scanner G2505B and Feature Extraction software (v10.1)
|
| Description |
Biological replicate 3 of 3
|
| Data processing |
The raw fluorescence intensity data were processed using the Polyphemus™ software (Oryzon Genomics), Q-Splines normalization.
|
| |
|
| Submission date |
Jul 27, 2009 |
| Last update date |
Oct 02, 2009 |
| Contact name |
angela tingaud-sequeira |
| E-mail(s) |
angela.tingaud@u-bordeaux1.fr
|
| Organization name |
Université Bordeaux 1
|
| Lab |
GPP
|
| Street address |
avenue des Facultés
|
| City |
Talence |
| ZIP/Postal code |
33400 |
| Country |
France |
| |
|
| Platform ID |
GPL8931 |
| Series (2) |
| GSE17333 |
Oogenesis in Solea senegalensis: Vitellogenic versus mature ovaries |
| GSE17337 |
Oogenesis in Solea senegalensis: various developmental stages |
|
