GSM5412667: -hsf1/msn2Δ/msn4Δ 42°C 30min Replicate3; Saccharomyces cerevisiae; RNA-Seq
Total RNA was extracted using a modified protocol based on Collart and Oliviero, 1993. Briefly, shock frozen cells were suspended in TES buffer and acidic phenol was added.  The suspension was vigorously shaken for 30 min at 65°C. The total RNA was extracted by phase separation with the classic phenol-chloroform procedure. Libraries were prepared with the SENSE mRNA-Seq Library Prep Kit V2 (Lexogen, Cat. No. 001.96) according to the manufacturers instructions. Briefly, total RNA was denatured for 2 min at 60 °C and polyA RNA was captured. After reverse transcription and second strand synthesis libraries were amplified and quality assessment was performed. The finished libraries were sequenced on a Illumina HiSeq1500 as 50 bp single reads with an average sequencing depth of 15 million reads per sample.