GSM4002758: NEU_P46; Homo sapiens; RNA-Seq
Total RNA was extracted with Quick-RNA MiniPrep Kit (Zymo Research – catalog number R1054). RNA quality was assessed through the RNA 6000 Nano Kit (Agilent) on an Agilent Bioanalyzer. RNA Integrity Number (RIN) was determined for every sample. Only samples with RIN >7.5 were processed. Sequencing libraries were prepared in two batches using TruSeq Stranded mRNA Library Prep (Illumina). 800 nanograms of total RNA were used as input. Oligo-attached magnetic beads were used for removing ribosomal RNA and globin mRNA from RNA samples. After a purification step, samples RNA were fragmented at 94°C for 8 minutes. RNA fragments were retro-transcribed to cDNA using Random Hexamer Primers. All cDNAs were indexed and amplified with 11 PCR cycles. Final libraries were validated with the Agilent DNA 1000 kit (Agilent) and sequenced on a NextSeq 500 platform (Illumina), producing 75 × 2 bp paired-end reads.