|
| Status |
Public on Mar 02, 2011 |
| Title |
WholeCell_HeminFeeding_60min_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Whole cells
|
| Organism |
Chlamydomonas reinhardtii |
| Characteristics |
strain: CC-124
|
| Treatment protocol |
For feeding experiments, a starter culture grown in the light with ~2 × 106 cells per ml was subdivided into 25 ml aliquots in 100 ml Erlenmeyer flasks and incubation continued in the dark for 16 h. At time 0, MgProto or hemin (final concentration 9 μM each) was added to the cultures in the dark and incubation in the dark was continued until harvest. Control cultures were treated the same way but did not receive the tetrapyrroles.
|
| Growth protocol |
Strains were grown heterotrophically in Tris-Acetate-Phosphate (TAP) medium (Harris, 1989) on a rotary shaker at 23°C either in the dark or under continuous irradiation with white light (fluence rate 45 μEm-2s-1) provided by fluorescent tubes (Osram L36W/25).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
As described in von Gromoff et al., 1989, Mol Cell Biol 9: 3911-3918
|
| Label |
Cy3
|
| Label protocol |
0.5 µg of total RNA were reverse transcribed into cDNA, and Cy3 labeled using the Quick Amp Labeling Kit, one-color (Agilent 5190-0442)
|
| |
|
| Hybridization protocol |
Hybridization was done with 0.6 µg labeled cDNA per array according to the Agilent protocol for 8x15k single color microarrays at a temperature of 65˚C overnight.
|
| Scan protocol |
An Agilent G2565CA DNA microarray scanner was used to scan arrays at 5 μm resolution, Feature Extraction Software 9.5.3.1 was then used to process and analyse array images.
|
| Description |
n/a
|
| Data processing |
The raw data was analyzed using GeneSpring GX 11.0 (Normalization: Shift to 75.0 percentile; Baseline Transformation: Median of all samples).
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| |
|
| Submission date |
Mar 12, 2010 |
| Last update date |
Mar 02, 2011 |
| Contact name |
Bjoern Voss |
| Organization name |
University of Freiburg
|
| Department |
Institute of Biology III
|
| Lab |
Genetics
|
| Street address |
Schänzlestr. 1
|
| City |
Freiburg |
| ZIP/Postal code |
79104 |
| Country |
Germany |
| |
|
| Platform ID |
GPL10173 |
| Series (2) |
| GSE20860 |
Change in gene expression in Chlamydomonas reinhardtii upon feeding with hemin and Mg-protoporphyrin |
| GSE20861 |
Change in gene expression in Chlamydomonas reinhardtii upon heat shock and feeding with hemin and Mg-protoporphyrin |
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