Ion Torrent Proton sequencing; RNA-seq of high lipid producing Yarrowia lipolytica during nitrogen or carbon restriction (control strain)
RNA-seq of high lipid producing Yarrowia lipolytica during nitrogen or carbon restriction (control strain)
Samples for RNA analysis were rapidly taken from steady-state chemostats, frozen in liquid nitrogen and stored at −80°C. Chemostat cultures were performed with a dilution rate of 0.05 h–1 at 30 °C in 1.2 L bioreactors (DASGIP) with a working volume of 750 mL at pH 3.5, controlled with 2 M KOH. Dissolved oxygen was kept above 30% with a stirrer rate of 600 rpm and an air flow rate of 1 vvm. For carbon restriction, one liter medium contained 5 g glucose, 5 g (NH4)2SO4, 3 g KH2PO4, 0.5 g MgSO4·7 H2O, vitamins and trace metal solutions (Verduyn et al. 1992) and 125 μL antifoam 204 (Sigma-Aldrich). For nitrogen restriction a similar medium was used, where the glucose was increased to 25 g, the (NH4)2SO4 decreased to 0.5 g while SO4 levels remained constant by adding 5.96 g K2SO4. CO2 and residual O2 in the exhaust gas were measured using an online gas analyser (DASGIP). Total RNA was extracted with Trizol (Invitrogen) using the manufacturer’s instructions with additional mechanical disruption of the cells using a FastPrep homogenizer (MP Biomedicals) and 1 mm silica beads.  Samples were depleted of rRNA using RiboZero
DEPARTMENT OF BIOLOGY AND BIOLOGICAL ENGINEERING, CHALMERS UNIVERSITY OF TECHNOLOGY
E-MTAB-5284:Sample 5_s
Department of Biology and Biological Engineering, Chalmers University of Technology
E-MTAB-5284:Sample 5_s
Experimental Factor: carbon restriction
growth condition