Dataset features


Application: Gene expression microarray analysis
Number of samples: 8
Release date: Mar 1 2007
Last update date: Mar 17 2012
Access: Public
Chemicals: Glucose, Glycerol
Dataset link Profiling of Rgt1-regulated gene transcripts (30º C) in the fungal pathogen Candida albicans

Experimental Protocol

A C. albicans genespecific microarray consisting of 70-mer oligonucleotides representing 6,346 of the 6,354 predicted ORFs in the annotated C. albicans genome assembly 19 was used. Each ORF was represented by a specific oligonucleotide, and one genome equivalent of this oligo was spotted three times per slide. To identify Rgt1-regulated transcripts, C. albicans cells from the wild type strain BWP17 or the rgt1 mutant strain CM18 were cultured in synthetic medium containing 5% glycerol at 30º C. Cells were harvested and RNA was extracted, labeled, and hybridized to the arrays. Each experiment consisted of a pair-wise competetive hybridization of total RNA samples derived from the wild type and mutant strains and included a reciprocal dye-flip replicate. Since two biological duplicates were tested for each strain, a total of four DNA microarray slides were used. Slides were scanned immediately after hybridization on a ScanArray express HT scanner (Perkin-Elmer). The scanner photomultiplier tube (PMT) values were set for optimal intensity with minimal background. An additional scan was done for each slide (low PMT) with the PMT such that <1% of the elements were saturated in order to characterise spots which were saturated at the higher PMT setting.