|
| Status |
Public on Nov 04, 2019 |
| Title |
S. cerevisiae_BY4741_SO2 60min_ biological rep2 |
| Sample type |
RNA |
| |
|
| Source name |
S. cerevisiae_BY4741_SO2 60min
|
| Organism |
Saccharomyces cerevisiae |
| Characteristics |
strain: BY4741 (MATa, his3-delta-1, leu2-delta-0, met15-delta-0, ura3-delta-0)
|
| Treatment protocol |
Cells of S. cerevisiae BY4741 and of the mutant BY4741_∆com2 were cultivated in MMB growth medium (at pH 3.5) in the absence (Control) or presence (Treatment) of sulphur dioxide.
|
| Growth protocol |
Cells of S. cerevisiae BY4741 and of the mutant BY4741_∆com2 were cultivated in MMB growth medium (at pH 3.5)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA isolation was performed using the hot-phenol method (Koher & Domdey, 1991, Methods in Enzimology 194:398-405).
|
| Label |
biotin
|
| Label protocol |
RNA was processed for use on Affymetrix (Santa Clara, CA, USA) GeneChip Yeast Genome 2.0 Arrays, according to the manufacturer's GeneChip 3' IVT Express kit user manual. Briefly, 100 ng of total RNA containing spiked in Poly-A RNA controls was used in a reverse transcription reaction (GeneChip 3' IVT Express Kit; Affymetrix) to generate first-strand cDNA. After second-strand synthesis, double-stranded cDNA was used in a 16h in vitro transcription (IVT) reaction to generate aRNA (GeneChip 3' IVT Express Kit; Affymetrix). Size distribution of the aRNA and fragmented aRNA, respectively, was assessed using an Agilent 2100 Bioanalyzer with a RNA 6000 Nano Assay.
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| |
|
| Hybridization protocol |
5 µg of fragmented aRNA was used in a 100-µl hybridization cocktail containing added hybridization controls. 80 µl of mixture was hybridized on arrays for 16 h at 45°C. Standard post hybridization wash and double-stain protocols (FS450_0003; GeneChip HWS kit, Affymetrix) were used on an Affymetrix GeneChip Fluidics Station 450.
|
| Scan protocol |
Arrays were scanned on an Affymetrix GeneChip scanner 3000 7G.
|
| Description |
Gene expression data from cells incubated for 60 min in the presence of sulphur dioxide
|
| Data processing |
MAS 5.0
|
| |
|
| Submission date |
Jul 30, 2018 |
| Last update date |
Nov 04, 2019 |
| Contact name |
Ana Mendes-Ferreira |
| E-mail(s) |
anamf@utad.pt
|
| Phone |
+351259350550
|
| Organization name |
Universidade de Trás-os-Montes e Alto Douro
|
| Street address |
Quinta de Prados
|
| City |
Vila Real |
| ZIP/Postal code |
5000-801 |
| Country |
Portugal |
| |
|
| Platform ID |
GPL2529 |
| Series (1) |
| GSE117883 |
The Msn2/4-like gene transcription factor Com2 (ORF YER130) is a key player of S. cerevisiae response and tolerance to sulphur dioxide |
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