title: NextSeq 500 paired end sequencing; RNA sequencing experiment of S. Cerevisiae strain CEN.PK113-7D in four different chemostat conditions
design: RNA sequencing experiment of S. Cerevisiae strain CEN.PK113-7D in four different chemostat conditions
library_name: Ana_Rep2_p
library_construction_protocol: 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.
attributes.tag: Experimental Factor: environmental stress
attributes.value: anaerobic condition; low glucose