organism: Yarrowia lipolytica
description: Protocols: 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
attributes.tag: ENA first public
attributes.value: 2017-06-07
attributes.tag: ENA last update
attributes.value: 2016-11-30
attributes.tag: External Id
attributes.value: SAMEA17802418
attributes.tag: INSDC center alias
attributes.value: Department of Biology and Biological Engineering, Chalmers University of Technology
attributes.tag: INSDC center name
attributes.value: Department of Biology and Biological Engineering, Chalmers University of Technology
attributes.tag: INSDC first public
attributes.value: 2017-06-07T17:03:12Z
attributes.tag: INSDC last update
attributes.value: 2016-11-30T09:36:02Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-5284:Sample 3
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: Po1g
attributes.tag: genetic modification
attributes.value: MATa, leu2-270, ura3-302::URA3, xpr2-332, axp-2 TEFin-LacZ-LEU2
attributes.tag: genotype
attributes.value: control strain
attributes.tag: growth condition
attributes.value: nitrogen restriction
attributes.tag: sample name
attributes.value: E-MTAB-5284:Sample 3