Illumina HiSeq 2000 paired end sequencing; RNA-Seq data of 16 strains of saccharomyces cerevisiae produced after knockout of all combination of HIS3, LEU2, URA3 and MET15 genes
RNA-Seq data of 16 strains of saccharomyces cerevisiae produced after knockout of all combination of HIS3, LEU2, URA3 and MET15 genes
S. cerevisiae strains were transferred from cryo-preserved cultures to agar dishes containing drop-in media and grown for 2 days at 30C. Pre-cultures were inoculated in 10 ml drop-in media and grown for 30 h at 30C. The cultures were washed with H2O and the main-cultures (n=5) inoculated at 0.15 OD600 in 200 l synthetic complete (SC) media in a 96-well plate. Growth was recorded for 35 hour for every 14 min by measuring OD595 using a FLUOstar OPTIMA (BMG Labtech) plate reader (87 rpm, 30C). Main-cultures were inoculated in 50 ml synthetic complete media at a starting OD600 of 0.15 (30C, 180 rpm). Strains were then grown for 6-19 hour until an OD600 of 0.8, the cells collected by centrifugation (2 min, 3000 g) and the supernatant discarded. The pellet was re-suspended in 1 ml H2O and three aliquots prepared with 350 l each. The cells were collected (0.5 min, 5000 g), the supernatant removed with a pipette and the sample snap frozen and stored at 80C until further processing. RNA for transcriptomics was prepared using the yeast RNA mini kit (Zymo Research) followed by DNase treatment, and processed using the TruSeq RNA library preparation kit (Illumina) following the manufacturers instructions