|
| Status |
Public on May 07, 2010 |
| Title |
Spermatogenesis in S. senegalensis: Mid vs. Late 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Mid spermatogenic testis
|
| Organism |
Solea senegalensis |
| Characteristics |
tissue: testis pieces
developmental stage: mid spermatogenesis
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from mid spermatogenic testes 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
|
| |
|
| Channel 2 |
| Source name |
Late spermatogenic testis
|
| Organism |
Solea senegalensis |
| Characteristics |
tissue: testis pieces
developmental stage: late spermatogenesis
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from late spermatogenic testes 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
|
| |
|
| |
| 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 2 of 3
|
| Data processing |
Raw data were obtained using Agilent's DNA Microarray Scanner G2505B and Feature Extraction software (v10.1). The raw fluorescence intensity data were processed using the Polyphemus software, developed at Oryzon Genomics, which includes spatial data compensation, non-significant expressed data filtering, and data normalization. Data normalization was carried out by an improved version of the nonlinear Q-splines normalization method, enhanced with robust regression techniques.
|
| |
|
| Submission date |
May 03, 2010 |
| Last update date |
May 06, 2010 |
| Contact name |
Josep V Planas |
| E-mail(s) |
jplanas@ub.edu
|
| Phone |
+34-93-4039384
|
| Organization name |
Universitat de Barcelona
|
| Department |
Fisiologia i Immunologia (Biologia)
|
| Lab |
Fisiologia Molecular de Peixos
|
| Street address |
Av. Diagonal 643
|
| City |
Barcelona |
| ZIP/Postal code |
08028 |
| Country |
Spain |
| |
|
| Platform ID |
GPL8931 |
| Series (2) |
| GSE21637 |
Transcriptional profiling of flatfish (Solea senegalensis) spermatogenesis: mid versus late spermatogenesis |
| GSE21694 |
Transcriptional profiling of flatfish (Solea senegalensis) spermatogenesis |
|
