BY4716xCBS432 Hybrid Citrinin Treated Replicate 2
A diploid hybrid yeast strain was produced by mating Sc (BY4716: MATα lys2 ura3∷KAN) and Sp (CBS432: MATa ura3∷HYG). Four single colony picks of the hybrid strain were grown overnight in 3 ml cultures of YPD medium at 30°C in a C24 Incubator Shaker (New Brunswick Scientific). Upon confirming that the cultures were in log phase, two of the replicates were treated with 91 µl of DMSO, while the other two were treated with 91 µl of DMSO containing citrinin such that its final concentration in the culture was 600 ppm. Cultures were allowed to continue shaking at 30°C for two additional hours followed by harvesting of the pellets by centrifugation.  RNA was immediately isolated from the pellets using the MasterPure™ Yeast RNA Purification Kit (Epicenter). Two µg of RNA from each of the samples was subsequently used to create sequencing libraries using the low-throughput protocol in the TruSeq RNA Sample Preparation Kit v2 (Illumina). Individually barcoded libraries were pooled and sequenced (single-end 101 bp reads) on a single lane of an Illumina HiSeq 2000 instrument at the Stanford Center for Genomics and Personalized Medicine.