NextSeq 500 sequencing; RNA-seq in wild-type Saccharomyces cerevisiae S288c cells to determine the effect of sodium crotonate treatment on the transcriptome
RNA-seq in wild-type Saccharomyces cerevisiae S288c cells to determine the effect of sodium crotonate treatment on the transcriptome
Three independent yeast colonies were used for inoculation. Overnight cultures were diluted in fresh medium with or without 10 mM sodium crotonate (Sigma-Aldrich) in the presence of 0.8 M sorbitol (Sigma-Aldrich). After 3.5 hours cells were in early log-phase (1 × 10^7 cells/ml) and harvested by centrifugation.  Total RNA was isolated using the RNeasyMini kit (Qiagen) and treated with the RNase-Free DNase Set (Qiagen) to remove any contaminating genomic DNA. Library preparations were performed with the TruSeq Stranded mRNA Library kit (Illumina).