RNAseq of homo sapiens: WT_CHOP-FmN_ISRIB_DMSO_2
RNA of 300 l TRIzollysate was isolated using the Maxwell RSC miRNA Tissue Kit (Promega, AS1460) following manufactures instructions. Afterwards, RNA concentration was quantified (NanoDrop, ThermoFisher) and 1 g of isolated RNA was additionally purified using Agencourt RNAClean XP Beads (Beckman Coulter, A63987). The purified RNA was quantified and quality controlled (Bioanalyzer, Agilent). For generating sequencing libraries 500 ng high quality RNA (RIN >8.0) were processed using the SENSE mRNA-Seq Lib Prep Kit V2 (Lexogen, A01161).  Data processing was done on Galaxy server hosted by LAFUGA, Gene Center, Munich. Raw sequencing data (fastq files)  was demultiplexed using Illumina Demultiplex v1.0.0. FastqFilter (Galaxy Version 1.0) was used to trim the data following instructions of Library Prep Kit (SENSE mRNA-Seq Lib Prep Kit V2 (Lexogen, A01161)). Trimmed reads were mapped against human genome (hg19) using STAR (Galaxy Version 2.5.2b-0). HTSEq-count (Galaxy Version 1.0.0) was used to analyze the abundancy of mapped reads to specific feature types (here: exon, minimum alignment quality 0, mode ontersection-strict, single-end, strand specificity reverse).