GSM1846308: yAS418_wt_RNASeq_alpha_control; Saccharomyces cerevisiae; RNA-Seq
~5 ODs of cells were used for RNA extraction. The cells were  spun down, flash frozen and stored at –80°C. The cells were thawed on ice, resuspended in 600µl RLT buffer with 6µl of beta-mercaptoethanol and added to 600µl beads. The cells were vortexed for 6 cycles (with 1 minute on, 1 minute off). The QIAGEN RNeasy Mini Kit purification of total RNA from yeast protocol was used to isolate the RNA. Total RNA quality and quantity were analyzed on an Agilent Bioanalyzer 2100 using a nano chip. RNAseq libraries were prepared using the Illumina TruSeq RNA sample prep v2 kit, starting from 500ng of total RNA, following the manufacturer's protocol with 13 PCR cycles. The amplified libraries were purified using AMPure beads, quantified by Qubit 2.0 fluorometer, and visualized on the Agilent Tapestation 2200. The libraries were pooled equimolarly, and loaded on the HiSeq 2500.