|
| Status |
Public on Jan 31, 2011 |
| Title |
C404A mutant, LoaOOH, rep1 |
| Sample type |
RNA |
| |
|
| Source name |
C404A mutant LoaOOH
|
| Organism |
Saccharomyces cerevisiae |
| Characteristics |
genotype: Swi6 C404A mutation
treatment: LoaOOH
|
| Treatment protocol |
Equal aliquots for linoleic acid hydroperoxide (LoaOOH) treatment (30 μM) for 45 min.
|
| Growth protocol |
Cultures were grown in 200 ml of SC-Leu to OD600 of 0.4 at 30°C .
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cultures were collected in tubes containing 40 g of –80°C crushed ice and cells pelleted by centrifugation (4000 g, 3 min), washed once in PBS before storage at –80°C. Frozen cell pellets were resuspended in 1 ml of Trizol® reagent (Invitrogen), lysed in a bead-beater with acid-washed glass beads and RNA was extracted as described by the manufacturer.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 6 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 ug of cRNA were processed through second-cycle single-stranded cDNA. Fragmented and labelled cDNA were then hybridized for 16 hr at 45C on an Affymetrix Yeast 2.0 array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
Gene arrays were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
CT6
Gene expression from C404A mutant LoaOOH rep1.
|
| Data processing |
Gene array data is processd by Expression Console software in which mean probeset signals for all probesets are analysed. A receiver operating curve (ROC) is generated by thresholding the computed value for the probesets. PLIER signals that pass this QC control is used for subsequent analysis.
|
| |
|
| Submission date |
Sep 29, 2009 |
| Last update date |
Jan 31, 2011 |
| Contact name |
Ruby CY Lin |
| E-mail(s) |
rubyl@unsw.edu.au, ruby.lin@sydney.edu.au
|
| Organization name |
The Westmead Institute for Medical Research
|
| Street address |
176 Hawkesbury Road, Westmead
|
| City |
Sydney |
| State/province |
NSW |
| ZIP/Postal code |
2145 |
| Country |
Australia |
| |
|
| Platform ID |
GPL2529 |
| Series (1) |
| GSE18334 |
Profiling oxidative stress in Saccharomyces cerevisiae by oxidation of transcription factor Swi6p |
|
