E-MTAB-6722:Glu_Rep3_p
NextSeq 500 paired end sequencing; RNA sequencing experiment of S. Cerevisiae strain CEN.PK113-7D in four different chemostat conditions
RNA sequencing experiment of S. Cerevisiae strain CEN.PK113-7D in four different chemostat conditions
Glu_Rep3_p
10OD of cells were collected, pelleted and snap frozen in liquid nitrogen. The S. cerevisiae strain CEN.PK113-7D was pre-cultured in a batch culture in Delft media with 2% glucose at 30°C and 200 rpm in shake flasks and then transferred to the chemostats in duplicates. Four different media compositions with a limitation in a different nutrient were employed in the chemostat runs. The temperature was also set to 30°C. The dilution rate was fixed to 0.1 and the cells were grown for 4 days to achieve stable cell numbers in the culture. Each environmental stress condition is described as follows: Low Glucose: minimal media with 0.75 % glucose and 1.35 g/l NH4. Anaerobic condition, low Glucose: minimal media with 1 % glucose and 1.35 g/l NH4, continuous gassed with N2. Low Ethanol: minimal media with 0.5 % ethanol and 1.35 g/l NH4. Low Nitrogen: minimal media with 6 % glucose and 0.27 g/l NH4 RNA was extracted using the RNeasy® Mini Kit from QIAGEN. Cells were mechanically disrupted using a FastPrep®-24 from MPbio in combination with the lysing matrix tubes type C from MPbio. The FastPrep was run 3 times for 20 seconds with 4.0m/s settings and a 5 minute break in between each run. RNA quality was assessed using NanoDrop and Bioanalyzer. The Illumina Kit TrueSeq RNA stranded HT kit was used following the manufacturers protocol.
Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, SE-41296, Sweden
E-MTAB-6722:Glu_Rep3_p
Experimental Factor: environmental stress
low glucose