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| Status |
Public on Dec 22, 2010 |
| Title |
Gis2 overexpression versus control cells |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Expression profiling by array
|
| Summary |
Measure the relative changes of gene expression upon GIS2 overexpression; 100 ml of BY4741 cells bearing plasmid pBG1805-Gis2 (GIS2 under the control of the galactose promotor) or the empty plasmid pBG1805 (=control) were grown in synthetic medium lacking uracil (SC-Ura supplemented with 2% raffinose) at 30 degrees to an OD600 of 0.45- 0.5. Expression of GIS2 was induced with 2% galactose for 1.5 h and cells were harvested by centrifugation, washed twice with 800 ul of ice-cold sterile water. 100 ul (1/8) of cells were removed and RNA was isolated by hot phenol extraction for microarray analysis. 6.7 ug of total RNA derived from cells expressing the empty vector (pBG1805) or Gis2p (pBG1805-Gis) were reverse transcribed in the presence of 5-(3-aminoallyl)-dUTP and natural dNTPs with a mixture of N9 and dT20V primers, and cDNAs were covalently linked to Cy3 and Cy5 NHS-monoesters (GE HealthSciences Cat# RPN5661), respectively, and competitively hybridized on yeast oligo arrays at 42 degrees celsius for 14 hours in formamide-based hybridization buffer. Microarrays were scanned with an Axonscanner 4200 (Molecular Devices) and analyzed with GenePix Pro 5.1 (Molecular devices).
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| Overall design |
Biological Replicate
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| |
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| Contributor(s) |
Scherrer T |
| Citation(s) |
21232131 |
| |
| Submission date |
Sep 07, 2010 |
| Last update date |
Mar 22, 2012 |
| Contact name |
Andre Gerber |
| E-mail(s) |
[email protected]
|
| Phone |
+41 44 6337378
|
| Fax |
+41 44 6337364
|
| Organization name |
Swiss Federal Institute of Technology
|
| Department |
Institute of Pharmaceutical Science
|
| Street address |
Wolfgang Pauli Str. 10, HCI H396
|
| City |
ETH Hoenggerberg, Zuerich |
| ZIP/Postal code |
ZH |
| Country |
Switzerland |
| |
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| Platforms (1) |
| GPL8546 |
SMD Print_1548 Saccharomyces cerevisiae |
|
| Samples (4)
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| This SubSeries is part of SuperSeries: |
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| Relations |
| BioProject |
PRJNA133211 |
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