title: GSM5225471: Experiment2_HOG1_KO_salt_3; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: Total RNA extraction was done with standard hot-phenol method. RNA integrity (RIN=8) was assessed by Bioanalyzer to ensure a RIN>8. Experiment 1: A total of 1μg of RNA was used for library preparation using the TruSeq stranded mRNA v2 (Illuimna) following manufacturer instructions. Quality control of generated libraries was performed with bioanalyzer and library quantification was done using the KAP kit (KapaBiosystems) prior to sequencing on an Illumina HiSeq2500 with 50 bp single-end reads; Experiment 2: A total of 5 μg of total RNA were used to purify mRNA using the polyA RNA selection magnetic beads following the manufacturer's instructions (Lexogen SKU: 039:100). Purified mRNA was used to generate strand-specific libraries using the total CORALL RNA kit (Lexogen SKU:  095.24). The number of library amplification cycles was determined by qPCR using a 1:10 dilution (PCR Add-on Lexogen, SKU: 020.96). Libraries were quantified by Qubit and Bioanalyzer. RNA-seq was performed on a pool containing all samples and triplicates and sequenced with 50 bp single-end read using an Illumina HiSeq2500.