Transcriptomics

Dataset Information

2

Comparison between Saccharomyces cerevisiae HS 10, HS 34 and HS 12, HS 37


ABSTRACT: Gene expression of different flocculent (HS 10 and HS 34) and powdery (HS 12 and HS 37) yeast strains compared to each other during exponential and stationary growth phase was analysed. The isolation of RNA was done by disruption of the cells under liquid nitrogen using mortar and pistil and then the Qiagen RNeasy Midi Kit with some modifications within the manufacturers´ protocol and the Qiagen RNase-Free DNase Set were applied. 6 µg of the total RNA per sample was used for each microarray experiments. The indirect labelling by the tyramide-signal-amplification method (MicromaxTM TSATM labelling and detection Kit from Perkin Elmer life sciences) was used to increase the Cy3 and Cy5 signals of microarray detection. Each cDNA containing Biotin- and Fluorescein-nucleotides respectively was purified with a QIAquick PCR purification kit and suspended in 11 µl of the formamide containing hybridization buffer. The slides were hybridized at 42°C over night under a cover slip. The microarrays were scanned by the Axon 4000B scanner; image intensity data were extracted and analysed with GenePix® Pro 6.0 software. Data from different scans of Dye-swap experiment were extracted by GenePix Pro 6.0 software, normalized and united. An outliertest has been applied in order to find outliers amongst the gene replicats. Subsequently, a t-test (1% and 5% probability of error) has been used in order to find regulated genes. Keywords: sorted yeast cells Microarray experiments were realised with dye swap.

ORGANISM(S): Saccharomyces cerevisiae  

SUBMITTER: Frank Schmidt   Cornelia Repenning  Dirk Benndorf  Claudia Wiacek  Franziska Heine  Thomas Scheper  Hauke Harms  Heike Sträuber  Frank Stahl  Martin von Bergen  Susann Müller 

PROVIDER: E-GEOD-12369 | ArrayExpress | 2008-12-11

SECONDARY ACCESSION(S): GSE12369PRJNA113201

REPOSITORIES: GEO, ArrayExpress

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Publications

Prediction of flocculation ability of brewing yeast inoculates by flow cytometry, proteome analysis, and mRNA profiling.

Heine Franziska F   Stahl Frank F   Sträuber Heike H   Wiacek Claudia C   Benndorf Dirk D   Repenning Cornelia C   Schmidt Frank F   Scheper Thomas T   von Bergen Martin M   Harms Hauke H   Müller Susann S  

Cytometry. Part A : the journal of the International Society for Analytical Cytology 20090201 2


The ability of brewing yeast to flocculate is an important feature for brewing of qualitatively good beer. Flocculation involves two main cell wall structures, which are the flocculation proteins (flocculins) and mannans, to which these flocculins bind. Unfortunately, in practice, the flocculation ability may get lost after several repitches. Flow cytometry was employed to analyze glucose and mannose structures of the cell surface by application of fluorescent lectins. Validation of the expressi  ...[more]

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