RNAseq of S. cerevisiae W303-GFP: control without OPP
Pre-inoculum cultures of Saccharomyces cerevisiae W303-GFP (control strain) were cultured at 30 C for 48 hours in 1 L baffle flasks containing 100 mL YNB w/o AA (pH 6.0; Difco) supplemented with the required amino acids and raffinose (2 %, w/v). Cells were collected via centrifugation (Eppendorf, F-45-12-11) for 5 min at 12 000 x g, washed twice in distilled water, re-suspended in 100 mL fresh YNB w/o AA (pH6.0) supplemented with the required amino acids and galactose (2 %, w/v). A total of 20 mL of cell suspension was incubated in 250 mL baffle flasks at 30 C for 4 hours. Cells were collected from a total of 4 mL culture volume via centrifugation (1 min; 12000 x g), followed by immediate removal of the supernatant and freezing of the pellet in liquid nitrogen. Preparation of total RNA was achieved using the Qiagen RNeasy Mini Kit (Qiagen, 74104) as per manufacturers instructions. All experiments were done in duplicate during five independent repeats (n = 10)
ScW303-GFP