GSM6909206: BV173 cells, sgPRDX1, untreated, 12h, biol rep 2; Homo sapiens; RNA-Seq
Total RNA was alcohol extracted from the cells resuspended in Trizol (Invitrogen) following manufacturer's protocols. RNA sequencing was performed using Illumina® Stranded mRNA Prep, Ligation (Illumina,  20040532) according to the manufacturers' protocol. The messenger RNA was captured form 1 μg of total RNA, fragmented and primed for cDNA synthesis followed by the second strand synthesis resulting in double-stranded cDNA. Secondly, pre-index anchors were ligated to the ends of the cDNA fragments, in the subsequent PCR amplification step the indexes (IDT® for Illumina® RNA UD Indexes Set A, Illumina, 20040553) and primer sequences for cluster generation were added. In order to optimize cluster densities across every lane of the flow cell, quantification of DNA libraries using DNA 1000 chip on an Agilent Technologies 2200 TapeStation System was performed. Indexed DNA libraries were normalized to 4 nM and then pooled in equal volumes. The library was sequenced on a Next Seq 550 system (Illumina) using NextSeq® Reagent Kit v3 (150 cycles) with a TG NextSeq 500/550 High Output Flow Cell. Images from the instrument were processed to generate FASTQ sequence files.