|
| Status |
Public on Apr 20, 2010 |
| Title |
Kinetic 3, Time point 21H day 1, technical replicates 1-3 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Ostreococcus tauri strain isolated from the Thau lagoon; 12:12 light/dark cycles (kinetic point)
|
| Organism |
Ostreococcus tauri |
| Characteristics |
time: 21H, day 1
strain: OTTH95
protocol: LIGHT OFF
|
| Treatment protocol |
LIGHT OFF
|
| Growth protocol |
Growth in Keller medium at 25°C under light/dark cycles
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted with RNeasy Plant Mini Kit (Qiagen) following manufacturer's instructions
|
| Label |
CY5
|
| Label protocol |
Total RNAs (350 ng) were amplified and labeled using a two-color labeling protocol Low Input Linear RNA amplification kit (Agilent) according to the manufacturer recommendations . Test and reference samples were respectively labeled with Cyanine-5 and Cyanine-3 CTP (10 mM, Perkin-Elmer/NENLife Science)
|
| |
|
| Channel 2 |
| Source name |
Ostreococcus tauri strain isolated from the Thau lagoon (reference)
|
| Organism |
Ostreococcus tauri |
| Characteristics |
strain: OTTH95
reference: pooled samples (3 kinetics)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted with RNeasy Plant Mini Kit (Qiagen) following manufacturer's instructions
|
| Label |
CY3
|
| Label protocol |
Total RNAs (350 ng) were amplified and labeled using a two-color labeling protocol Low Input Linear RNA amplification kit (Agilent) according to the manufacturer recommendations . Test and reference samples were respectively labeled with Cyanine-5 and Cyanine-3 CTP (10 mM, Perkin-Elmer/NENLife Science)
|
| |
|
| |
| Hybridization protocol |
Hybridization was performed using Agilent In Situ Hybridization Kit Plus. 1 microg of each test and reference cRNAs were mixed and subjected to fragmentation (30 min at 60°C). Samples were diluted in Nexterion Hyb buffer (Schott Nexterion) and hybridized using Gasket Slides in an Agilent Hybridization rotation oven (60°C, 17 h at 4 rpm in the dark). Slides were disassembled, washed according to the Schott protocol
|
| Scan protocol |
Scanned with the dynamic autofocus Agilent G2565BA microarray scanner.
Images were quantified using Agilent Feature Extraction Software (version 9.1).
|
| Description |
Biological replicate 3 (kinetic 3 of 3)
|
| Data processing |
Normalization was performed using the print-tip loess method and scaled with the Gquantile method
The value of each biological time point is calculate as an average of the technical triplicate.
|
| |
|
| Submission date |
Jun 03, 2009 |
| Last update date |
Apr 20, 2010 |
| Contact name |
annabelle monnier |
| E-mail(s) |
annabelle.monnier@univ-rennes1.fr
|
| Phone |
+33 (0)2 23 23 61 14
|
| Fax |
+33 (0)2 23 23 44 78
|
| URL |
http://www.umr6061.univ-rennes1.fr
|
| Organization name |
UMR 6061 Génétique et Développement
|
| Street address |
2 avenue du Professeur Léon Bernard
|
| City |
Rennes |
| ZIP/Postal code |
35043 |
| Country |
France |
| |
|
| Platform ID |
GPL8644 |
| Series (1) |
| GSE16422 |
Genome wide rhythmic transcription under light/dark cycles in Ostreococcus tauri |
|
