Culture was grown for 18 hours at 30 C in a rotating wall vessel placed on a rotator set at 30 rpm
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted using a combination of the hot phenol/glass beads method of Ausubel et al. (Ausubel FM, Brent E, Kingstion RE, Moore DD, Seidman JG, Smith JA, and Struhl K. Current Protocols in Molecular Biology, New York: Wiley, 1995) and TRIZOL (Sigma).
Label
Biotin
Label protocol
Total RNA (10 µg) was converted into double-stranded cDNA (ds-cDNA) by using SuperScript Choice System (Life Technologies, Rockville, MD) with an oligo(dT) primer containing a T7 RNA polymerase promoter (Genset, San Diego, CA). After second-strand synthesis, the reaction mixture was extracted with phenol-chloroform-isoamyl alcohol, and ds-cDNA was recovered by ethanol precipitation.
Hybridization protocol
Standard Affymetrix procedures
Scan protocol
Standard Affymetrix procedures
Description
Saccharomyces cerevisiae strain BY4743 gorwn in a rotating wall vessel (RWV) for 18 hours on a rotator
