|
| Status |
Public on Feb 01, 2014 |
| Title |
Transformant yeast 10 µg/ml fludioxonil 15 min rep2 |
| Sample type |
RNA |
| |
|
| Source name |
S. cerevisiae expressing C. albicans Nik1p, fludioxonil, 15 min
|
| Organism |
Saccharomyces cerevisiae |
| Characteristics |
strain: BWG1-7a
expression vector: pYES2+CaNIK1
treatment: 10 µg/ml fludioxonil
duration of treatment: 15 min
|
| Treatment protocol |
Induction of transgene expression was achieved by incubation of transformants in SG-ura at 30°C for 16h, and functional analysis of the recombinant strain was carried out in SG-ura.
Cultures were treated with 10 µg/ml fludioxonil.
|
| Growth protocol |
S. cerevisiae transformants were maintained in SD-ura (0.67% yeast nitrogen base without amino acids, 2% glucose, nucleosides and amino acids according to Rose, M. D., Winston, F., and Hieter, P. (1990) Methods in Yeast Genetics: A Laboratory Course Manual, Revised Ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, without uracil) at 30°C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were disrupted using glass beads (425-600 µm, Sigma) in the presence of lysis buffer (Qiagen RLT). RNA was isolated using RNeasy columns (Qiagen) according to the manufacturer's protocol.
|
| Label |
biotin
|
| Label protocol |
500 ng of total RNA were used for biotin labeling according to the 3´ IVT Express Kit (Affymetrix).
|
| |
|
| Hybridization protocol |
Samples were hybridized to an identical lot of Affymetrix GeneChip Yeast Genome 2.0 Arrays for 16 hours at 45°C. Steps for washing and SA-PE staining were processed on the fluidics station 450 using the mini_euk2v3_FS450 protocol (Affymetrix).
|
| Scan protocol |
Image analysis was performed using a GCS3000 Scanner and GCOS1.2 Software Suite (Affymetrix).
|
| Description |
F15.2
|
| Data processing |
Raw intensities were RMA (robust multi-array average) normalized using the affy package of Bioconductor/R.
|
| |
|
| Submission date |
Oct 28, 2010 |
| Last update date |
Jan 23, 2015 |
| Contact name |
Anna Buschart |
| E-mail(s) |
[email protected]
|
| Organization name |
Helmholtz-Centre for Infection Research
|
| Department |
BiSA
|
| Street address |
Inhoffenstr. 7
|
| City |
Braunschweig |
| ZIP/Postal code |
38124 |
| Country |
Germany |
| |
|
| Platform ID |
GPL2529 |
| Series (1) |
| GSE25002 |
Expression data from Saccharomyces cerevisiae expressing Candida albicans Nik1p comparing osmotic stress and fludioxonil treatment |
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