Transcriptomics

Dataset Information

3

Gis2 overexpression versus control cells


ABSTRACT: 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). Biological Replicate

ORGANISM(S): Saccharomyces cerevisiae  

SUBMITTER: Andre Gerber   Tanja Scherrer 

PROVIDER: E-GEOD-24002 | ArrayExpress | 2010-12-22

SECONDARY ACCESSION(S): GSE24002PRJNA133211

REPOSITORIES: GEO, ArrayExpress

Similar Datasets

2010-06-24 | E-GEOD-15297 | ArrayExpress
2008-07-18 | E-GEOD-11152 | ArrayExpress
2010-06-25 | E-GEOD-15376 | ArrayExpress
2008-07-29 | E-GEOD-12094 | ArrayExpress
2010-06-23 | E-GEOD-16252 | ArrayExpress
2007-11-05 | E-GEOD-9046 | ArrayExpress
2007-08-15 | E-GEOD-8770 | ArrayExpress
2007-08-17 | E-GEOD-7466 | ArrayExpress
2010-09-09 | E-GEOD-9047 | ArrayExpress
2010-06-25 | E-GEOD-6797 | ArrayExpress