GSM8718602: RPE1-TP53KO-EIF2AKO biorep 3 flow cell 2; Homo sapiens; RNA-Seq
RNeasy Mini kit (Qiagen) according to manufacturer's instructions. 25 * 106 paired-end reads of 75 bp per sample from a NovaSeq 6000 were aligned to the Hg19 genome build using STAR, to obtain gene-level counts The NEBNext Ultra II Directional RNA Library Prep Kit for Illumina was used to process the sample(s). The sample preparation was performed according to the protocol "NEBNext Ultra II Directional RNA Library Prep Kit for Illumina" (NEB #E7760S/L). Briefly, mRNA was isolated from total RNA using the oligo-dT magnetic beads. After fragmentation of the mRNA, a cDNA synthesis was performed. This was used for ligation with the sequencing adapters and PCR amplicification of the resulting product. The quality and yield after sample preparation was measured with the Fragment Analyzer (see appendices). The size of the resulting products was consistent with the expected size distribution (a broad peak between 300-500 bp).